Optimization of living cell delivery parameters in air-assisted atomization

空气辅助雾化的细胞递送参数的优化 Optimization of living cell delivery parameters in air-assisted atomization 1 2 first-author 严心涛（1987-），男，湖北仙桃人，硕士，副研究员，2009年于西南交通大学获得学士学位，2011年于哈尔滨工业大学获得硕士学位，主要从事生物医学测量方法研究及仪器设计开发工作。E-mail： yanxt@sibet.ac.cn. http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996242&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996248&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996245&type= 22.01333237 32.00400162 严心涛 （1987-），男，湖北仙桃人，硕士，副研究员，2009年于西南交通大学获得学士学位，2011年于哈尔滨工业大学获得硕士学位，主要从事生物医学测量方法研究及仪器设计开发工作。E-mail： yanxt@sibet.ac.cn . yanxt@sibet.ac.cn 2 2 2 1 2 corresp E-mail： wuxiaodong2000@hotmail.com E-mail： wuxiaodong2000@hotmail.com 武晓东（1968-），男，吉林舒兰县人，博士生导师，研究员，现任中国科学院苏州生物医学工程技术研究所副所长，江苏省医用光学重点实验室主任，主要从事医用激光技术、光学工程、医用光学等方面研究工作。E-mail： wuxiaodong2000@hotmail.com http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996254&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996259&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996256&type= 22.01333618 32.00399780 武晓东 （1968-），男，吉林舒兰县人，博士生导师，研究员，现任中国科学院苏州生物医学工程技术研究所副所长，江苏省医用光学重点实验室主任，主要从事医用激光技术、光学工程、医用光学等方面研究工作。E-mail： wuxiaodong2000@hotmail.com wuxiaodong2000@hotmail.com 复旦大学 工程与应用技术研究院，上海 200433 复旦大学 工程与应用技术研究院，上海 200433 Academy for Engineering & Technology， Fudan University， Shanghai 200433， China Academy for Engineering & Technology， Fudan University， Shanghai 200433， China 中国科学院 苏州生物医学工程技术研究所 中科院生物医学检验技术重点实验室， 江苏 苏州 215163 中国科学院 苏州生物医学工程技术研究所 中科院生物医学检验技术重点实验室， 江苏 苏州 215163 CAS Key Lab of Bio-Medical Diagnostics， Suzhou Institute of Biomedical Engineering and Technology， Chinese Academy of Sciences， Suzhou 215163， China CAS Key Lab of Bio-Medical Diagnostics， Suzhou Institute of Biomedical Engineering and Technology， Chinese Academy of Sciences， Suzhou 215163， China 为实现生物墨水的高效率递送，构建了一种基于空气辅助雾化技术的细胞递送装置，并利用多种方法系统地测量了雾化特性，以及雾化参数对细胞活性的影响。根据粒径的Rosin-Rammler分布规律对雾化均匀度指数进行了定量评估；利用多帧图像叠加以及局部阈值二值化方法对雾化视频进行边界提取，分析雾化角；结合BM3D去噪算法与二值化算法精确定位雾滴位置，实现雾滴运动速度的精确测量。实验结果表明：雾化高度和射流流量均与雾滴粒径正相关；当辅助气体压力高于60 kPa时，会显著影响雾滴均匀度；通过射流流量和雾化高度的参数控制，雾化面积可实现50~1 800 mm 2 的大宽幅调节；雾滴的运动速度受雾化高度影响较大，且在50 mm雾化高度下运动速度最大，均值可高达14 m/s；雾化高度和辅助空气流量均会显著影响HaCaT细胞活性，在50 mm雾化高度下细胞活化率低至64.01±0.86%（阴性对照组为92.98±3.21%），而在100 mm雾化高度下可高达90.24±0.73%；HaCaT细胞活性在喷涂后72小时内与未喷涂的保持一致。该雾化装置可用于生物墨水的高效率递送，为实现不同面积喷涂或细胞高活化率递送的雾化参数优化设计提供了指导。 To achieve high-efficiency delivery of bio-ink， a novel cell delivery device based on air-assisted atomization was developed，and its atomization characteristics and their effect on cell viability were studied. According to the Rosin-Rammler droplet size distribution law， the uniformity index was quantitatively evaluated. Multi-frame image superposition and local threshold binarization were used to extract the boundary of atomization from a video， so that the atomization angle could be analyzed. By combining the BM3D denoising and binarization algorithm to accurately locate the droplet positions， we could calculate the velocity of the droplets. The atomization height and the liquid flow rate are positively correlated to the diameter of the droplets. The pressure of the assistant air has a noticeable effect on the droplet uniformity when this pressure is higher than 60 kPa. By controlling the liquid flow rate and spray height， the spray area can be adjusted to a wide range of 50-1 800 mm 2 . The velocity of droplets was greatly affected by the spray height. At a spray height of 50 mm， the velocity reached maximum value， with an average value as high as 14 m/s. Both the spray height and the auxiliary air flow rate significantly affect the viability of HaCaT cells. The rate of cell activation was 64.01±0.86% and 90.24±0.73% at a spray height of 50 mm and 100 mm， respectively （the cell activation rate was 92.98±3.21% in the negative control group）. Within 72 hours after spraying， the viability of HaCaT cells was consistent with that of unsprayed cells. The novel atomization device can be used for high-efficiency delivery of bio-ink， and the relevant research results also provide guidance for the optimal design of atomization parameters to achieve spraying of areas of different size and high cell-activation rate delivery. 空气辅助雾化 雾化 细胞递送 雾化特性 雾滴速度 细胞存活率 air-assisted atomization atomization cell delivery spray characteristic droplet velocity cell viability 1 引 言 1 大面积皮肤伤口与多种突发事件、重大疾病紧密相关，是临床上常见病症，其迁延不愈的特性严重降低了患者的生活质量，甚至危及生命，并且在全球范围内造成了巨大的经济负担 ［ 1 1 - 2 2 1-2 ］ 。由大面积皮肤创伤而引发的一系列严重并发症和感染是威胁患者生命的两个重要因素 ［ 3 3 ］ 。因此，除了给患者静脉注射液体和营养素以抵消脱水和补充失去的蛋白质外，采取有效的治疗方法以减轻患者疼痛和挽回生命显得至关重要。目前临床实践的有效治疗方法主要为自体皮肤移植术 ［ 4 4 - 5 5 4-5 ］ ，但对于自体皮源本已稀缺的患者，该方法仍面临着巨大挑战。 细胞移植术是解决这个问题的有效方法 ［ 6 6 - 8 8 6-8 ］ ，即从患者身上采集自体皮肤细胞（通常是角质形成细胞）后直接递送到伤口，或经过细胞培养传代后悬浮，再递送到伤口。事实证明，细胞移植术可以显著影响细胞在伤口中的分布和增殖 ［ 9 9 - 11 11 9-11 ］ 。近些年，细胞移植术已经从静脉注射、肌肉注射、注射器或移液枪的局部滴/注 ［ 12 12 - 13 13 12-13 ］ 发展到雾化式喷洒 ［ 14 14 - 16 16 14-16 ］ 。尽管静脉注射、肌肉注射或局部细胞给药均显示出治疗效果，但前两种给药方法会导致作用于伤口部位的细胞浓度低，治疗效果不佳 ［ 12 12 ］ ；通过移液枪或注射器进行局部细胞给药虽可减少患者疼痛，但细胞递送效率低，且分布不均匀 ［ 13 13 ］ 。细胞悬浮液喷雾自体移植用自体表皮衍生细胞，使用更小的自体部位（供体部位与烧伤面积比例 ε 可达1∶100）即可在短时间内完成再上皮化 ［ 16 16 - 17 17 16-17 ］ ，对于深部、大面积和不规则创面，该方法能实现细胞的快速和高效的递送。目前涉及组织修复与再生应用的空气喷枪或雾化装置仍存在一些缺陷，如生成的雾滴大小不一致、分布不均匀 ［ 18 18 ］ ，雾化参数对雾化特性的精确控制尚不清楚 ［ 9 9 ］ ，在辅助空气压力为69 kPa下，细胞活化率仅为65% ［ 19 19 ］ ，有相关文献报道 ［ 20 20 ］ 对细胞雾化递送4天后，细胞增殖能力大受影响，细胞活化率降为45%。如何实现药物的均匀化雾化、不同面积的雾化或较高的细胞活化率雾化是细胞给药雾化递送装置开发中需面临的几个关键技术问题。 空气辅助雾化方法具有仅需较低空气压力和射流压力即可实现较好的雾化效果，以及较大的射流通道可避免堵塞等特点 ［ 21 21 ］ ，本文构建了基于该雾化方法的可用于细胞递送的雾化装置，通过对其雾化特性测试方法研究，侧重分析雾化高度、辅助气体流量和射流流量等雾化参数对雾化特性和生物墨水（Bio-ink）递送的影响，以指导雾化参数的优化设计和提高生物墨水的雾化喷涂效率。 2 装置设计及试剂材料 1 2.1　装置原理及设计 2 空气辅助雾化的细胞递送装置（Air-Assisted Atomization Device，AAAD）的工作原理如 图1 图1 （a）所示，空气泵输出的气体通过空气滤芯过滤后分两个流路，一路经气压调节阀后在雾化喷嘴出口处形成稳定的高速涡旋气流，另一路也经气压调节阀以稳定压力驱动无菌储液管中细胞悬浮液，并在喷嘴的针管出口形成液柱或液膜。在喷嘴出口处，气体与生物墨水之间会形成很高的相对速度，从而破坏液柱或液膜，改变生物墨水的流动性、表观粘性和表面张力，最终使连续相的生物墨水变成分散相以达到雾化效果。其中，两个流路中气压调节阀分别用于调节辅助空气气压以控制气体流量和调节无菌储液管的驱动压力以控制射流流量。为分析雾化高度对雾化性能的影响，雾化喷嘴可沿导杆做上下滑动。 AAAD主要包含气动控制模块和雾化喷嘴模块两部分，实物分别如 图1 图1 （b）和 图1 图1 （c）所示。装置主要组成部件包含流量为10 L/min的微型空气压缩泵（中国，SKOOCOM），100 kPa与200 kPa量程且灵敏度均优于0.2%的精密调压阀（日本，SMC），储液筒（日本，MUSASHI），27G雾化针管（日本，MUSASHI），雾化针管的内径为200~250 μm，喷嘴的内径为0.7~1.0 mm。采用量程为5 L/min且精度优于1.5%的微型空气流量计（中国，SENLOD）和量程为40 mL/min且精度优于5%的微型液体流量计（瑞士，SENSIRION）来测量辅助气体流量和射流流量。 2.2　试剂材料 2 人永生化表皮细胞（HaCaT，中国，COBIOER）用含10%胎牛血清（FBS，中国，TIANHANG）和1%青霉素/链霉素（A5955，Sigma）的高糖培养基（DMEM，Gibco）在T75 cm培养瓶（美国，CORNING）中培养，孵育条件为37 ℃、CO 2 的体积分数为5%。每4天传代一次，每隔一天更换一次培养基。当细胞达到90%以上汇合时，用胰蛋白酶（15090046，Gibco）收集细胞，将细胞悬液在室温下以1 000 r/min的转速离心5 min，弃去上清液，再将细胞重悬于无菌生理盐水（ST341，中国，Beyotime）中，配制成细胞密度约为10 6 cells/mL的生物墨水。在雾化基本特性测试中，均以无菌生理盐水为研究对象。 3 雾化特性测试方法 1 3.1　雾滴粒径分布及均匀度分析 2 本文采用激光粒度仪（JL-3000，中国，JNGX）对雾滴粒径进行测量。该仪器主要基于夫琅和费（Fraunhofer）衍射理论测量雾滴尺寸及对应的雾滴个数。雾化喷嘴产生的雾滴是由大小不等的雾滴群组成，本文雾滴粒径分布采用体积累积分布表示，平均粒径采用体积平均粒径 D ［4，3］计算，即： <math display="block" id="M1"><mi>D</mi><mo stretchy="false">[</mo><mn mathvariant="normal">4,3</mn><mo stretchy="false">]</mo><mo>=</mo><mfrac><mrow><mo>∑</mo><mi>n</mi><mo>⋅</mo><msup><mrow><mi>D</mi></mrow><mrow><mn mathvariant="normal">4</mn></mrow></msup></mrow><mrow><mo>∑</mo><mi>n</mi><mo>⋅</mo><msup><mrow><mi>D</mi></mrow><mrow><mn mathvariant="normal">3</mn></mrow></msup></mrow></mfrac></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996277&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996274&type= 30.90333366 10.92199993 . 经换算可得： <math display="block" id="M2"><mi>D</mi><mo stretchy="false">[</mo><mn mathvariant="normal">4,3</mn><mo stretchy="false">]</mo><mo>=</mo><mo>∑</mo><mi>f</mi><mo>⋅</mo><mi>D</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996282&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996280&type= 28.19400024 4.74133301 ， 其中： <math id="M3"><mi>n</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996287&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996285&type= 1.86266661 2.79399991 是各个单一粒径对应的雾滴个数； <math id="M4"><mi>D</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996294&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996290&type= 2.79399991 2.79399991 是雾滴的粒径，单位为μm； <math id="M5"><mi>f</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996540&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996537&type= 1.10066664 3.64066648 是各单一粒径的雾滴群对应的体积频度。 通常，雾滴粒径是不连续的，但当雾滴数量足够多时，可假设雾滴的粒径累计分布是连续的，通常其符合Rosin-Rammler分布（简称“R-R”分布）规律 ［ 22 22 ］ ，即： <math display="block" id="M6"><mi>F</mi><mo stretchy="false">(</mo><msub><mrow><mi>D</mi></mrow><mrow><mi>i</mi></mrow></msub><mo stretchy="false">)</mo><mo>=</mo><mn mathvariant="normal">1</mn><mo>-</mo><mi mathvariant="normal">e</mi><mi mathvariant="normal">x</mi><mi mathvariant="normal">p</mi><mtext> </mtext><mfenced open="[" close="]" separators="|"><mrow><mo>-</mo><msup><mrow><mfenced separators="|"><mrow><mfrac><mrow><msub><mrow><mi>D</mi></mrow><mrow><mi>i</mi></mrow></msub></mrow><mrow><mover accent="true"><mi>D</mi><mo>¯</mo></mover></mrow></mfrac></mrow></mfenced></mrow><mrow><mi>k</mi></mrow></msup></mrow></mfenced></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996305&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996303&type= 42.75666428 11.34533310 ， 其中： <math id="M7"><mi>F</mi><mo stretchy="false">(</mo><msub><mrow><mi>D</mi></mrow><mrow><mi>i</mi></mrow></msub><mo stretchy="false">)</mo></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996312&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996309&type= 9.90600014 4.48733330 表示雾滴粒径小于 <math id="M8"><msub><mrow><mi>D</mi></mrow><mrow><mi>i</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996319&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996315&type= 3.55599999 3.72533321 的体积累计百分比； <math id="M9"><mover accent="true"><mi>D</mi><mo>¯</mo></mover></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996328&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996324&type= 2.79399991 3.30200005 为雾滴的特征尺寸，即 <math id="M10"><mi>F</mi><mo stretchy="false">(</mo><msub><mrow><mi>D</mi></mrow><mrow><mi>i</mi></mrow></msub><mo stretchy="false">)</mo><mo>=</mo><mn mathvariant="normal">0.632</mn></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996334&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996331&type= 23.02933502 4.48733330 对应的雾滴粒径； <math id="M11"><mi>k</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996345&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996343&type= 1.69333339 2.79399991 为均匀度指数， <math id="M12"><mi>k</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996345&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996343&type= 1.69333339 2.79399991 越大，表示雾滴粒径分布的均匀性越好。对于符合“R-R”分布规律的雾化，若粒径 <math id="M13"><msub><mrow><mi>D</mi></mrow><mrow><mi>i</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996351&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996349&type= 3.55599999 3.72533321 和 <math id="M14"><msub><mrow><mi>D</mi></mrow><mrow><mi>j</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996358&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996355&type= 3.55599999 3.72533321 对应的累计百分比分别为 <math id="M15"><mi>F</mi><mo stretchy="false">(</mo><msub><mrow><mi>D</mi></mrow><mrow><mi>i</mi></mrow></msub><mo stretchy="false">)</mo></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996363&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996361&type= 9.90600014 4.48733330 和 <math id="M16"><mi>F</mi><mo stretchy="false">(</mo><msub><mrow><mi>D</mi></mrow><mrow><mi>j</mi></mrow></msub><mo stretchy="false">)</mo></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996369&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996366&type= 9.90600014 4.48733330 ，则由 式（3） 式（3） 不难推导出雾化均匀度指数 <math id="M17"><mi>k</mi><mo stretchy="false">[</mo><msub><mrow><mi>D</mi></mrow><mrow><mi>i</mi></mrow></msub><mo>,</mo><msub><mrow><mi>D</mi></mrow><mrow><mi>j</mi></mrow></msub><mo stretchy="false">]</mo></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996377&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996373&type= 15.40933418 4.48733330 ，即： <math display="block" id="M18"><mi>k</mi><mo stretchy="false">[</mo><msub><mrow><mi>D</mi></mrow><mrow><mi>i</mi></mrow></msub><mo>,</mo><msub><mrow><mi>D</mi></mrow><mrow><mi>j</mi></mrow></msub><mo stretchy="false">]</mo><mo>=</mo><mfrac><mrow><mn mathvariant="normal">1</mn></mrow><mrow><mi mathvariant="normal">l</mi><mi mathvariant="normal">g</mi><mtext> </mtext><msub><mrow><mi>D</mi></mrow><mrow><mi>i</mi></mrow></msub><mo>-</mo><mi mathvariant="normal">l</mi><mi mathvariant="normal">g</mi><msub><mrow><mi>D</mi></mrow><mrow><mi>j</mi></mrow></msub></mrow></mfrac><mfenced separators="|"><mrow><mi mathvariant="normal">l</mi><mi mathvariant="normal">g</mi><mi mathvariant="normal">l</mi><mi mathvariant="normal">n</mi><mtext> </mtext><mfrac><mrow><mn mathvariant="normal">1</mn></mrow><mrow><mn mathvariant="normal">1</mn><mo>-</mo><mi>F</mi><mo stretchy="false">(</mo><msub><mrow><mi>D</mi></mrow><mrow><mi>i</mi></mrow></msub><mo stretchy="false">)</mo></mrow></mfrac><mo>-</mo><mi mathvariant="normal">l</mi><mi mathvariant="normal">g</mi><mi mathvariant="normal">l</mi><mi mathvariant="normal">n</mi><mtext> </mtext><mfrac><mrow><mn mathvariant="normal">1</mn></mrow><mrow><mn mathvariant="normal">1</mn><mo>-</mo><mi>F</mi><mo stretchy="false">(</mo><msub><mrow><mi>D</mi></mrow><mrow><mi>j</mi></mrow></msub><mo stretchy="false">)</mo></mrow></mfrac></mrow></mfenced><mo>.</mo></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996382&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996379&type= 98.72133636 10.49866676 3.2　雾化角测试及雾化面积评估 2 雾化过程中雾滴群在空间呈圆锥状分布，该圆锥的张角即为雾化角，其大小直接影响雾化面积 A 。本文采用工业相机（DMK 23G445，德国，THE IMAGING SOURCE）对雾化状态摄像后，在Matlab（美国，MathWorks）中计算雾化角。使用宽工作距离范围的物镜（BLADE 2.4/12， Korea， FIM optcis），摄像分辨率为640×480像素，其中每个像素对应物方尺寸为139 μm。 由于单帧图像信噪比（Signal-to-Noise Ratio，SNR）较低，边界提取不准确，而随着图像叠加数量的增加，SNR也将增大。经测试，单帧图像的SNR仅为11.8 dB，用于雾化边界提取误差较大，通过对多帧图像进行叠加，可显著提高图像的SNR。当叠加图像达到227帧时，图像SNR为50 dB，对叠加后的高SNR图像进行二值化处理，能够有效保留雾化边界，且边界更加清晰。随着图像的叠加数量增多，SNR将进一步增大，如当叠加视频中所有图像时（总帧数为238），图像SNR可达65.5 dB。本文计算雾化角的方法是：先对雾化过程进行连续视频拍摄，分析时，叠加视频的所有帧的图像，以提高图像SNR，如 图2 图2 （a）为单帧图像， 图2 图2 （b）为视频叠加图像；对雾化角图像进行二值化，采用局部自适应阈值分割算法，该算法可根据局部光强分布自动调整二值化阈值，对于边界的识别更加准确和稳定，如 图2 图2 （c）； 图2 图2 （d）为二值化图像的边界数据点，考虑到靠近喷嘴和远离喷嘴处的雾化角度有所变化，因此数据点的分析也分为两部分，采用最小二乘法拟合数据点，得到边界直线方程，进而计算得到靠近喷嘴的雾化角 <math id="M19"><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">1</mn></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996389&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996385&type= 2.96333337 3.72533321 和远离喷嘴处的雾化角 <math id="M20"><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">2</mn></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996394&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996392&type= 2.96333337 3.72533321 。如 图2 图2 （d）所示，计算出雾化角 <math id="M21"><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">1</mn></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996400&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996397&type= 2.96333337 3.72533321 和 <math id="M22"><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">2</mn></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996821&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996819&type= 2.96333337 3.72533321 后，可进一步估算出生物墨水喷洒在基板上的面积 A ，即： <math display="block" id="M23"><mi>A</mi><mo>=</mo><mfenced open="{" close="" separators="|"><mrow><mtable><mtr><mtd><mi>π</mi><mo>⋅</mo><msup><mrow><mfenced open="[" close="]" separators="|"><mrow><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">N</mi></mrow></msub><mo stretchy="false">(</mo><mi mathvariant="normal">t</mi><mi mathvariant="normal">a</mi><mi mathvariant="normal">n</mi><mtext> </mtext><mfrac><mrow><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">1</mn></mrow></msub></mrow><mrow><mn mathvariant="normal">2</mn></mrow></mfrac><mo>-</mo><mi mathvariant="normal">t</mi><mi mathvariant="normal">a</mi><mi mathvariant="normal">n</mi><mtext> </mtext><mfrac><mrow><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">2</mn></mrow></msub></mrow><mrow><mn mathvariant="normal">2</mn></mrow></mfrac><mo stretchy="false">)</mo><mo>+</mo><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub><mo>⋅</mo><mi mathvariant="normal">t</mi><mi mathvariant="normal">a</mi><mi mathvariant="normal">n</mi><mtext> </mtext><mfrac><mrow><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">2</mn></mrow></msub></mrow><mrow><mn mathvariant="normal">2</mn></mrow></mfrac></mrow></mfenced></mrow><mrow><mn mathvariant="normal">2</mn></mrow></msup><mo>,</mo></mtd><mtd><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">N</mi></mrow></msub><mo>≤</mo><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></mtd></mtr><mtr><mtd><mi>π</mi><mo>⋅</mo><msup><mrow><mfenced separators="|"><mrow><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">N</mi></mrow></msub><mo>⋅</mo><mi mathvariant="normal">t</mi><mi mathvariant="normal">a</mi><mi mathvariant="normal">n</mi><mtext> </mtext><mfrac><mrow><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">1</mn></mrow></msub></mrow><mrow><mn mathvariant="normal">2</mn></mrow></mfrac></mrow></mfenced></mrow><mrow><mn mathvariant="normal">2</mn></mrow></msup><mo>,</mo></mtd><mtd><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">N</mi></mrow></msub><mo>></mo><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></mtd></mtr></mtable></mrow></mfenced></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996419&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996416&type= 85.68266296 22.69066620 ， 其中： <math id="M24"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">N</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996425&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996422&type= 3.55599999 3.72533321 为最小二乘法拟合直线断点位置距离喷嘴出口位置的高度， <math id="M25"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996431&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996428&type= 3.38666677 3.72533321 为雾化基板距离喷嘴出口位置的高度，单位均为mm。 3.3　雾滴沉降速度 2 由于雾滴沉降速度会影响细胞活性 ［ 23 23 ］ ，本文利用高速相机（HS1，德国，PCO）的双快门捕获功能来测算雾滴沉降速度，相机单次曝光时间设置为10 μs，拍摄时间间隔 T 为3.58 μs。采用10×远距离物镜（375-039，日本，三丰），视场为1.65×2.75 mm 2 。由于曝光时间极短，拍摄图像SNR较低，如 图3 图3 （a），很难对视场中的雾滴进行准确定位。本文采用BM3D去噪算法，可有效提高图像质量，如 图3 图3 （b）；利用二值化算法对图像中的雾滴进行识别后，如 图3 图3 （c），根据雾滴光强分布计算得到雾滴质心坐标 <math id="M26"><mfenced separators="|"><mrow><msub><mrow><mi>x</mi></mrow><mrow><mi>T</mi><mn mathvariant="normal">1</mn></mrow></msub><mo>,</mo><msub><mrow><mi>z</mi></mrow><mrow><mi>T</mi><mn mathvariant="normal">1</mn></mrow></msub></mrow></mfenced></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996436&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996434&type= 13.97000027 3.97933316 和 <math id="M27"><mfenced separators="|"><mrow><msub><mrow><mi>x</mi></mrow><mrow><mi>T</mi><mn mathvariant="normal">2</mn></mrow></msub><mo>,</mo><msub><mrow><mi>z</mi></mrow><mrow><mi>T</mi><mn mathvariant="normal">2</mn></mrow></msub></mrow></mfenced></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996442&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996439&type= 13.97000027 3.97933316 ，高速相机双快门捕获图像处理前与处理后分别如 图3 图3 （d）和 图3 图3 （e）所示；再根据相邻两帧图像中雾滴的坐标，分别计算出雾滴的横向运动速度 <math id="M28"><msub><mrow><mi>v</mi></mrow><mrow><mi>x</mi></mrow></msub><mo>=</mo><mrow><mrow><mfenced open="|" close="|" separators="|"><mrow><msub><mrow><mi>x</mi></mrow><mrow><mi>T</mi><mn mathvariant="normal">2</mn></mrow></msub><mo>-</mo><msub><mrow><mi>x</mi></mrow><mrow><mi>T</mi><mn mathvariant="normal">1</mn></mrow></msub></mrow></mfenced></mrow><mo>/</mo><mrow><mi>T</mi></mrow></mrow></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996447&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996445&type= 28.19400024 4.48733330 ，纵向运动速度 <math id="M29"><msub><mrow><mi>v</mi></mrow><mrow><mi>z</mi></mrow></msub><mo>=</mo><mrow><mrow><mfenced open="|" close="|" separators="|"><mrow><msub><mrow><mi>z</mi></mrow><mrow><mi>T</mi><mn mathvariant="normal">2</mn></mrow></msub><mo>-</mo><msub><mrow><mi>z</mi></mrow><mrow><mi>T</mi><mn mathvariant="normal">1</mn></mrow></msub></mrow></mfenced></mrow><mo>/</mo><mrow><mi>T</mi></mrow></mrow></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996453&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996450&type= 27.26266861 4.48733330 ，以及合速度 <math id="M30"><msub><mrow><mi>v</mi></mrow><mrow><mi mathvariant="normal">r</mi></mrow></msub><mo>=</mo><mroot><mrow><msup><mrow><msub><mrow><mi>v</mi></mrow><mrow><mi>x</mi></mrow></msub></mrow><mrow><mn mathvariant="normal">2</mn></mrow></msup><mo>+</mo><msup><mrow><msub><mrow><mi>v</mi></mrow><mrow><mi>z</mi></mrow></msub></mrow><mrow><mn mathvariant="normal">2</mn></mrow></msup></mrow><mrow/></mroot></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996458&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996455&type= 24.72266769 4.74133301 。 3.4　细胞活化率测试 2 将配制好的生物墨水一部分用于无喷涂的阴性对照，采用注射器（2 mL，中国，KINDLY）挤压递送；另一部分用AAAD喷洒。为了评估雾化后活细胞数与死细胞数比例，采用Calcein/PI细胞活性与细胞毒性检测试剂盒（C2015M，中国，Beyotime）在37 ℃避光孵育30 min，再用倒置荧光显微镜（IX73，日本，OLYMPUS）拍摄荧光图片，并利用ImageJ（美国，National Institutes of Health）对荧光图像进行自动计数和Merge处理。细胞活化率 <math id="M31"><mi>η</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996979&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996977&type= 1.86266661 3.64066648 由活细胞总数占细胞总数（含活细胞与死细胞）的百分比来表示。采用CCK-8试剂盒（C0043，中国，Beyotime）检测喷涂或未喷涂的细胞在24 h、48 h及72 h后的活性，来分析AAAD喷涂对细胞增殖性能的影响。 公开的文献［ 15 15 ］中已分析了辅助空气压力 <math id="M32"><msub><mrow><mi>P</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996504&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996502&type= 4.14866638 3.72533321 对 <math id="M33"><mi>η</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996979&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996977&type= 1.86266661 3.64066648 的影响，即在 <math id="M34"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996489&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996486&type= 3.38666677 3.72533321 为 <math id="M35"><mn mathvariant="normal">100</mn><mtext> </mtext><mi mathvariant="normal">m</mi><mi mathvariant="normal">m</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996494&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996492&type= 12.19200039 2.87866688 ， <math id="M36"><msub><mrow><mi>P</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub><mo>≤</mo><mn mathvariant="normal">70</mn><mtext> </mtext><mi mathvariant="normal">k</mi><mi mathvariant="normal">P</mi><mi mathvariant="normal">a</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996501&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996498&type= 19.30400085 4.23333359 时， <math id="M37"><msub><mrow><mi>P</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996504&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996502&type= 4.14866638 3.72533321 对 <math id="M38"><mi>η</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996979&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996977&type= 1.86266661 3.64066648 无显著差异。在本文中，重点分析 <math id="M39"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996579&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996576&type= 3.38666677 3.72533321 和辅助空气流量 <math id="M40"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996964&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996962&type= 4.57200003 4.14866638 对 <math id="M41"><mi>η</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996979&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996977&type= 1.86266661 3.64066648 的影响。 3.5　统计方法 2 数据表示为平均值±标准偏差（Standard Deviation，SD），并使用Origin（美国，OriginLab）做非配对 t 检验（Unpaired t test）、单因素方差分析（One-Way ANOVA）或双因素方差分析（Two-way ANOVA），以评估各实验组之间的差异性。 4 结果与讨论 1 4.1　射流流量和雾化高度对雾滴粒径分布的影响 2 图4 图4 为雾滴粒径分布及雾化参数对其影响。 <math id="M42"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996964&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996962&type= 4.57200003 4.14866638 设定为3 L/min，在不同 <math id="M43"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996828&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996827&type= 4.31799984 4.14866638 和不同 <math id="M44"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996944&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996942&type= 3.38666677 3.72533321 的雾化条件下，经激光粒度仪得到不同雾滴粒径及对应的体积频度 <math id="M45"><mi>f</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996540&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996537&type= 1.10066664 3.64066648 ，如 图4 图4 （a）所示，由 式（2） 式（2） 可计算出 D ［4，3］，根据 式（3） 式（3） 可以得到“R-R”分布曲线，在该曲线上分别选取25%和75%累计百分比对应下的粒径大小 D 25和 D 75，一般选取 D 25和 D 75之间的分布情况来说明颗粒的集中情况 ［ 15 15 ］ ，利用 式（4） 式（4） 计算出 <math id="M46"><mi>k</mi><mfenced open="[" close="]" separators="|"><mrow><mi>D</mi><mn mathvariant="normal">25</mn><mo>,</mo><mi>D</mi><mn mathvariant="normal">75</mn></mrow></mfenced></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996632&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996630&type= 20.23533440 3.97933316 来定性分析雾滴的均匀度程度。 通过双因素方差分析，可得出 <math id="M47"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996579&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996576&type= 3.38666677 3.72533321 、 <math id="M48"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996828&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996827&type= 4.31799984 4.14866638 以及两者间的交互作用均对 D ［4，3］有显著影响。利用单因素方差分析来具体探究它们之间的关系，如 图4 图4 （b），不难得到：（1）在同等 <math id="M49"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996944&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996942&type= 3.38666677 3.72533321 下， <math id="M50"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996828&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996827&type= 4.31799984 4.14866638 对 D ［4，3］有显著影响，表现为 <math id="M51"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996828&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996827&type= 4.31799984 4.14866638 越大， D ［4，3］越大；（2） <math id="M52"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub><mo>=</mo><mn mathvariant="normal">0.2</mn><mtext> </mtext><mi mathvariant="normal">m</mi><mi mathvariant="normal">L</mi><mo>/</mo><mi mathvariant="normal">m</mi><mi mathvariant="normal">i</mi><mi mathvariant="normal">n</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996575&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996573&type= 27.60133362 4.40266657 时， <math id="M53"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996579&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996576&type= 3.38666677 3.72533321 对 D ［4，3］无显著影响；（3）以高度为25 mm实验组为参考组，在相同 <math id="M54"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996584&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996580&type= 4.31799984 4.14866638 条件下，50 mm实验组与参考组差异不大，但75 mm实验组和100 mm实验组与参考组相比均显著增大，这说明在雾化过程中，当 <math id="M55"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996587&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996586&type= 3.38666677 3.72533321 大于一定值后，随着其进一步增大，雾滴与雾滴间存在融合现象，使得 D ［4，3］有显著增大趋势，但当 <math id="M56"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996828&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996827&type= 4.31799984 4.14866638 过低时， <math id="M57"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996944&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996942&type= 3.38666677 3.72533321 对 D ［4，3］无显著影响。 封装细胞的雾滴与基板发生撞击中可能造成细胞损失，一般而言， D ［4，3］越小，细胞活化率 <math id="M58"><mi>η</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996979&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996977&type= 1.86266661 3.64066648 越低 ［ 23 23 ］ ；但 D ［4，3］过大时，细胞位于雾滴边缘概率会增加，雾滴与基板撞击过程中，细胞受到的剪切应力会增大， <math id="M59"><mi>η</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996979&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996977&type= 1.86266661 3.64066648 降低 ［ 24 24 ］ 。因此， <math id="M60"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996608&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996606&type= 3.38666677 3.72533321 与 <math id="M61"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996828&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996827&type= 4.31799984 4.14866638 在细胞递送中是重要雾化参数。细胞传送装置在实际使用中 <math id="M62"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996828&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996827&type= 4.31799984 4.14866638 一般低于5 mL/min ［ 14 14 ， 16 16 ， 25 25 ］ ，否则会影响生物墨水的利用率。 经测量，当 <math id="M63"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub><mo>=</mo><mn mathvariant="normal">3</mn><mtext> </mtext><mi mathvariant="normal">L</mi><mo>/</mo><mi mathvariant="normal">m</mi><mi mathvariant="normal">i</mi><mi mathvariant="normal">n</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996620&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996618&type= 21.92866707 4.40266657 时， <math id="M64"><msub><mrow><mi>P</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub><mo>≈</mo><mn mathvariant="normal">20</mn><mtext> </mtext><mi mathvariant="normal">k</mi><mi mathvariant="normal">P</mi><mi mathvariant="normal">a</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996624&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996622&type= 19.30400085 3.80999994 。利用单因素方差分析可以得到：在H2实验组和H4实验组中， <math id="M65"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996828&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996827&type= 4.31799984 4.14866638 对 <math id="M66"><mi>k</mi><mfenced open="[" close="]" separators="|"><mrow><mi>D</mi><mn mathvariant="normal">25</mn><mo>,</mo><mi>D</mi><mn mathvariant="normal">75</mn></mrow></mfenced></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996632&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996630&type= 20.23533440 3.97933316 有显著不同的影响，但影响程度不一致，如 图4 图4 （c）所示，说明这两组实验的雾滴生成状态不稳定；但在H1~H4四组实验中，当 <math id="M67"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996636&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996634&type= 4.31799984 4.14866638 相同时， <math id="M68"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996639&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996637&type= 3.38666677 3.72533321 对 <math id="M69"><mi>k</mi><mfenced open="[" close="]" separators="|"><mrow><mi>D</mi><mn mathvariant="normal">25</mn><mo>,</mo><mi>D</mi><mn mathvariant="normal">75</mn></mrow></mfenced></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996671&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996669&type= 20.23533440 3.97933316 影响不大。为进一步分析喷嘴出口的辅助空气流场对雾化的影响，研究了20~100 kPa辅助气体压力下的雾滴分布状态，如 图4 图4 （d）所示，可得到：在 <math id="M70"><msub><mrow><mi>P</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub><mo>≤</mo><mn mathvariant="normal">50</mn><mtext> </mtext><mi mathvariant="normal">k</mi><mi mathvariant="normal">P</mi><mi mathvariant="normal">a</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996647&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996645&type= 19.55800056 4.23333359 时，各实验组的 <math id="M71"><mi>k</mi><mfenced open="[" close="]" separators="|"><mrow><mi>D</mi><mn mathvariant="normal">25</mn><mo>,</mo><mi>D</mi><mn mathvariant="normal">75</mn></mrow></mfenced></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996671&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996669&type= 20.23533440 3.97933316 相差不显著；而在 <math id="M72"><msub><mrow><mi>P</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub><mo>≥</mo><mn mathvariant="normal">60</mn><mtext> </mtext><mi mathvariant="normal">k</mi><mi mathvariant="normal">P</mi><mi mathvariant="normal">a</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996655&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996653&type= 19.55800056 4.14866638 时，各实验组的 <math id="M73"><mi>k</mi><mfenced open="[" close="]" separators="|"><mrow><mi>D</mi><mn mathvariant="normal">25</mn><mo>,</mo><mi>D</mi><mn mathvariant="normal">75</mn></mrow></mfenced></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996659&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996657&type= 20.23533440 3.97933316 显著提高，尤其在80 kPa和90 kPa实验组中，相对于前一个梯度实验组而言， <math id="M74"><mi>k</mi><mfenced open="[" close="]" separators="|"><mrow><mi>D</mi><mn mathvariant="normal">25</mn><mo>,</mo><mi>D</mi><mn mathvariant="normal">75</mn></mrow></mfenced></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996663&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996661&type= 20.23533440 3.97933316 提高地非常显著。虽然 <math id="M75"><msub><mrow><mi>P</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996667&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996665&type= 4.14866638 3.72533321 的增大能提高 <math id="M76"><mi>k</mi><mfenced open="[" close="]" separators="|"><mrow><mi>D</mi><mn mathvariant="normal">25</mn><mo>,</mo><mi>D</mi><mn mathvariant="normal">75</mn></mrow></mfenced></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996671&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996669&type= 20.23533440 3.97933316 ，但同样会影响细胞递送中 <math id="M77"><mi>η</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996679&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996677&type= 1.86266661 3.64066648 值，在文献［ 15 15 ］中已做了相关分析。因此，在一些生物材料需均匀化喷涂而不考虑 <math id="M78"><mi>η</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996679&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996677&type= 1.86266661 3.64066648 指标时，可以重点关注 <math id="M79"><msub><mrow><mi>P</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996683&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996681&type= 4.14866638 3.72533321 这项雾化参数。 4.2　雾化角及雾化面积的影响因素 2 图5 图5 为雾化参数对雾化角及雾化面积的影响。经最小二乘法拟合边界直线方程，在不同 <math id="M80"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996828&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996827&type= 4.31799984 4.14866638 下可计算得到 <math id="M81"><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">1</mn></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996817&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996815&type= 2.96333337 3.72533321 和 <math id="M82"><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">2</mn></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996821&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996819&type= 2.96333337 3.72533321 ，且 <math id="M83"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">N</mi></mrow></msub><mo>≈</mo><mn mathvariant="normal">25</mn><mtext> </mtext><mi mathvariant="normal">m</mi><mi mathvariant="normal">m</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996700&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996698&type= 18.79599953 3.80999994 。从 图5 图5 （a）中可看到：在 <math id="M84"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996703&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996702&type= 4.57200003 4.14866638 为 <math id="M85"><mn mathvariant="normal">3</mn><mtext> </mtext><mi mathvariant="normal">L</mi><mo>/</mo><mi mathvariant="normal">m</mi><mi mathvariant="normal">i</mi><mi mathvariant="normal">n</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996708&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996706&type= 12.61533356 3.30200005 的实验组中，随着 <math id="M86"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996828&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996827&type= 4.31799984 4.14866638 增大， <math id="M87"><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">1</mn></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996817&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996815&type= 2.96333337 3.72533321 先从26.44°增加到30.81°之后一直递减到8.04°，而 <math id="M88"><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">2</mn></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996718&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996716&type= 2.96333337 3.72533321 在15.55°~25.26°间上下波动；当 <math id="M89"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub><mo>≤</mo><mn mathvariant="normal">7</mn><mrow><mrow><mtext> </mtext><mi mathvariant="normal">m</mi><mi mathvariant="normal">L</mi></mrow><mo>/</mo><mrow><mi mathvariant="normal">m</mi><mi mathvariant="normal">i</mi><mi mathvariant="normal">n</mi></mrow></mrow></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996722&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996721&type= 24.63800049 3.64066648 时， <math id="M90"><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">1</mn></mrow></msub><mo>></mo><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">2</mn></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996727&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996725&type= 10.75266743 3.72533321 ；而在 <math id="M91"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996828&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996827&type= 4.31799984 4.14866638 为8 mL/min或9 mL/min时， <math id="M92"><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">1</mn></mrow></msub><mo><</mo><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">2</mn></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996734&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996732&type= 10.75266743 3.72533321 。结果表明，在 <math id="M93"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996738&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996736&type= 4.57200003 4.14866638 恒定条件下， <math id="M94"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996742&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996740&type= 4.31799984 4.14866638 对 <math id="M95"><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">1</mn></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996746&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996744&type= 2.96333337 3.72533321 的影响比对 <math id="M96"><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">2</mn></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996821&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996819&type= 2.96333337 3.72533321 的影响显著。利用 式（5） 式（5） 可计算出在不同 <math id="M97"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996828&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996827&type= 4.31799984 4.14866638 下的雾化面积 <math id="M98"><mi>A</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996778&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996776&type= 2.87866688 2.79399991 ，其中取 <math id="M99"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">N</mi></mrow></msub><mo>=</mo><mn mathvariant="normal">25</mn><mtext> </mtext><mi mathvariant="normal">m</mi><mi mathvariant="normal">m</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996763&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996760&type= 18.79599953 3.80999994 。不难发现在固定 <math id="M100"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996944&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996942&type= 3.38666677 3.72533321 下调节 <math id="M101"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996828&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996827&type= 4.31799984 4.14866638 时，在 <math id="M102"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub><mo>=</mo><mn mathvariant="normal">3</mn><mtext> </mtext><mi mathvariant="normal">m</mi><mi mathvariant="normal">L</mi><mo>/</mo><mi mathvariant="normal">m</mi><mi mathvariant="normal">i</mi><mi mathvariant="normal">n</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996774&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996772&type= 24.63800049 4.40266657 条件下的 <math id="M103"><mi>A</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996778&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996776&type= 2.87866688 2.79399991 最大，且 <math id="M104"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996944&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996942&type= 3.38666677 3.72533321 越大，差异越显著；在 <math id="M105"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub><mo>=</mo><mn mathvariant="normal">25</mn><mtext> </mtext><mi mathvariant="normal">m</mi><mi mathvariant="normal">m</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996786&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996784&type= 18.62666702 3.80999994 时，雾化面积可控制在100 mm 2 内，可满足小面积喷洒的工况，如 图5 图5 （b）所示。 如 图5 图5 （c），在 <math id="M106"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996796&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996794&type= 4.31799984 4.14866638 为为 <math id="M107"><mn mathvariant="normal">3</mn><mtext> </mtext><mrow><mrow><mi mathvariant="normal">m</mi><mi mathvariant="normal">L</mi></mrow><mo>/</mo><mrow><mi mathvariant="normal">m</mi><mi mathvariant="normal">i</mi><mi mathvariant="normal">n</mi></mrow></mrow></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996799&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996798&type= 14.90133286 3.21733332 的实验组中，当 <math id="M108"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub><mo>≤</mo><mn mathvariant="normal">1.5</mn><mtext> </mtext><mi mathvariant="normal">L</mi><mo>/</mo><mi mathvariant="normal">m</mi><mi mathvariant="normal">i</mi><mi mathvariant="normal">n</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996802&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996801&type= 24.89200020 4.40266657 时，雾化不充分，且 <math id="M109"><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">1</mn></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996817&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996815&type= 2.96333337 3.72533321 略小于 <math id="M110"><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">2</mn></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996821&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996819&type= 2.96333337 3.72533321 ；而当 <math id="M111"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub><mo>≥</mo><mn mathvariant="normal">2</mn><mtext> </mtext><mi mathvariant="normal">L</mi><mo>/</mo><mi mathvariant="normal">m</mi><mi mathvariant="normal">i</mi><mi mathvariant="normal">n</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996813&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996812&type= 21.92866707 4.40266657 时，雾化效果良好， <math id="M112"><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">1</mn></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996817&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996815&type= 2.96333337 3.72533321 和 <math id="M113"><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">2</mn></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996821&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996819&type= 2.96333337 3.72533321 会显著增大，且趋于平稳， <math id="M114"><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">1</mn></mrow></msub><mo>></mo><msub><mrow><mi>θ</mi></mrow><mrow><mn mathvariant="normal">2</mn></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996825&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996823&type= 10.75266743 3.72533321 。结果表明，在 <math id="M115"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996828&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996827&type= 4.31799984 4.14866638 恒定条件下，为形成良好的雾化效果， <math id="M116"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996964&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996962&type= 4.57200003 4.14866638 需足够大，建议不低于2 L/min。其中，部分雾化图像如 图5 图5 （d）所示。 4.3　气体流量和雾化高度对雾滴沉降速度的影响 2 图6 图6 （a）是雾滴沉降速度测试平台；在 <math id="M117"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">L</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996838&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996835&type= 4.31799984 4.14866638 为 <math id="M118"><mn mathvariant="normal">3</mn><mtext> </mtext><mi mathvariant="normal">m</mi><mi mathvariant="normal">L</mi><mo>/</mo><mi mathvariant="normal">m</mi><mi mathvariant="normal">i</mi><mi mathvariant="normal">n</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996841&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996839&type= 15.49400043 3.30200005 ， <math id="M119"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub><mo>≤</mo><mn mathvariant="normal">5</mn><mtext> </mtext><mi mathvariant="normal">L</mi><mo>/</mo><mi mathvariant="normal">m</mi><mi mathvariant="normal">i</mi><mi mathvariant="normal">n</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996844&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996843&type= 21.92866707 4.40266657 时，如 图6 图6 （b）所示，H1实验组的 <math id="M120"><msub><mrow><mi>v</mi></mrow><mrow><mi>x</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996878&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996876&type= 3.04800010 3.72533321 最高，随着雾化高度 <math id="M121"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996944&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996942&type= 3.38666677 3.72533321 变大， <math id="M122"><msub><mrow><mi>v</mi></mrow><mrow><mi>x</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996878&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996876&type= 3.04800010 3.72533321 会降低，但H3与H4实验组的 <math id="M123"><msub><mrow><mi>v</mi></mrow><mrow><mi>x</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996878&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996876&type= 3.04800010 3.72533321 变化不大；在各个实验组中， <math id="M124"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996964&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996962&type= 4.57200003 4.14866638 对 <math id="M125"><msub><mrow><mi>v</mi></mrow><mrow><mi>x</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996878&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996876&type= 3.04800010 3.72533321 影响不大；由 图6 图6 （c）和 图6 图6 （d）可知， <math id="M126"><msub><mrow><mi>v</mi></mrow><mrow><mi>z</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996895&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996893&type= 2.87866688 3.72533321 和 <math id="M127"><msub><mrow><mi>v</mi></mrow><mrow><mi mathvariant="normal">r</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996899&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996897&type= 2.70933342 3.72533321 均比 <math id="M128"><msub><mrow><mi>v</mi></mrow><mrow><mi>x</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996878&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996876&type= 3.04800010 3.72533321 大的多；在4个实验组中，H2实验组的 <math id="M129"><msub><mrow><mi>v</mi></mrow><mrow><mi>z</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996895&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996893&type= 2.87866688 3.72533321 和 <math id="M130"><msub><mrow><mi>v</mi></mrow><mrow><mi mathvariant="normal">r</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996899&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996897&type= 2.70933342 3.72533321 最大；在各个实验组中， <math id="M131"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996964&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996962&type= 4.57200003 4.14866638 对 <math id="M132"><msub><mrow><mi>v</mi></mrow><mrow><mi>z</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996895&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996893&type= 2.87866688 3.72533321 和 <math id="M133"><msub><mrow><mi>v</mi></mrow><mrow><mi mathvariant="normal">r</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996899&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996897&type= 2.70933342 3.72533321 影响也不显著。 结果表明，在辅助空气低流量下，雾滴的运动速度受 <math id="M134"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996909&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996907&type= 3.38666677 3.72533321 的影响较大；辅助气体流场主要在喷嘴出口的小范围内影响 <math id="M135"><msub><mrow><mi>v</mi></mrow><mrow><mi>x</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996913&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996911&type= 3.04800010 3.72533321 ，随着 <math id="M136"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996916&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996914&type= 3.38666677 3.72533321 变大，雾滴在空气阻力作用下， <math id="M137"><msub><mrow><mi>v</mi></mrow><mrow><mi>x</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996920&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996918&type= 3.04800010 3.72533321 会逐渐减小；雾滴的 <math id="M138"><msub><mrow><mi>v</mi></mrow><mrow><mi>z</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996924&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996922&type= 2.87866688 3.72533321 和 <math id="M139"><msub><mrow><mi>v</mi></mrow><mrow><mi mathvariant="normal">r</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996928&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996926&type= 2.70933342 3.72533321 在辅助气体的作用下，有一段加速积累的过程，需运行一定距离后才会逐渐降低。雾滴的运动速度直接影响其与雾化基板之间的剪切应力，为保证细胞雾化递送中的细胞活化率 <math id="M140"><mi>η</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996932&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996930&type= 1.86266661 3.64066648 ，雾滴的运动速度应尽可能低。因此，对于本文设计的雾化喷嘴，应避免在50 mm的雾化高度喷涂。 4.4　生物墨水雾化的细胞活化率影响因素 2 经倒置荧光显微镜拍摄和Image J软件分析后，可得出阴性对照组和不同 <math id="M141"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996944&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996942&type= 3.38666677 3.72533321 下实验组中 <math id="M142"><mi>η</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996979&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996977&type= 1.86266661 3.64066648 ，如 图7 图7 （a）。经单因素方差分析，得到 <math id="M143"><msub><mrow><mi>h</mi></mrow><mrow><mi mathvariant="normal">F</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996944&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996942&type= 3.38666677 3.72533321 对 <math id="M144"><mi>η</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996979&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996977&type= 1.86266661 3.64066648 有显著的影响（ p < 0.001）；将各个实验组与阴性对照组作非配对t检验，可得到在50 mm的雾化高度进行细胞传递时，确实会显著降低 <math id="M145"><mi>η</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996979&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996977&type= 1.86266661 3.64066648 （ p < 0.001），如 图7 图7 （b）所示，该结果同时证明了 <math id="M146"><mi>η</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996979&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996977&type= 1.86266661 3.64066648 与雾滴粒径分布和雾滴沉降速度均存在一定关联。 图7 图7 （c）结果表明，当 <math id="M147"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub><mo>≤</mo><mn mathvariant="normal">3</mn><mtext> </mtext><mi mathvariant="normal">L</mi><mo>/</mo><mi mathvariant="normal">m</mi><mi mathvariant="normal">i</mi><mi mathvariant="normal">n</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996960&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996958&type= 21.92866707 4.40266657 下， <math id="M148"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996964&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996962&type= 4.57200003 4.14866638 对 <math id="M149"><mi>η</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996979&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996977&type= 1.86266661 3.64066648 无显著影响；而在 <math id="M150"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub><mo>≥</mo><mn mathvariant="normal">4</mn><mtext> </mtext><mi mathvariant="normal">L</mi><mo>/</mo><mi mathvariant="normal">m</mi><mi mathvariant="normal">i</mi><mi mathvariant="normal">n</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996971&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996969&type= 21.92866707 4.40266657 下， <math id="M151"><msub><mrow><mi>Q</mi></mrow><mrow><mi mathvariant="normal">A</mi></mrow></msub></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996975&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996973&type= 4.57200003 4.14866638 对 <math id="M152"><mi>η</mi></math> http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996979&type= http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=32996977&type= 1.86266661 3.64066648 会有显著负作用。采用CCK-8试剂盒对HaCaT细胞在24 h、48 h以及72 h后增殖检测的结果显示，AAAD喷涂对细胞增殖性能无显著影响，如 图8 图8 所示。 5 结 论 1 本文构建了一种基于空气辅助雾化技术的可用于细胞递送的雾化装置；系统地研究了雾滴粒径分布、雾化角、雾滴沉降速度等雾化特性的光学测试方法、图像优化处理算法以及雾滴均匀度和雾化面积的定量评估方法；并利用统计学方法分析了多种雾化参数对雾化特性或细胞活化率的影响。结果表明：高于60 kPa的辅助气体压力与雾滴均匀度呈正相关关系，但会降低细胞活化率；通过控制射流流量和雾化高度，可实现50~1 800 mm 2 大宽幅的雾化面积调节；雾滴的运动速度受雾化高度影响较大，且在50 mm附近的运动速度最大，均值可达14 m/s；雾化高度显著影响着雾滴的粒径分布和运行速度，进而影响到细胞活化率，结果显示雾化高度为50 mm时对细胞活化率有非常显著影响，阴性对照细胞活化率均值为92.98%，在50 mm雾化高度时其仅为64.01%，而在100 mm雾化高度时其可高达90.24%。本文研制的雾化装置为细胞或生物材料提供了一种递送手段，雾化特性测试方法的研究也对雾化参数的优化设计和细胞或生物材料的喷涂效率提升具有指导意义。此外，雾化装置所需气源的输出流量不超过10 L/min即可实现较好的雾化效果，这为未来开发一体化手持式的细胞雾化喷枪创造了有利条件，在细胞移植术领域将具有广泛的应用前景。 参考文献 LI H S ， WANG S ， TAN J L ， et al . 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