皮肤组织扩张与细胞牵拉培养对表皮细胞生长及表达硫氧还蛋白影响的实验研究
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摘要
目的
建立充分模拟皮肤组织扩张过程的贴壁细胞机械牵拉培养体外模型；研究不同牵拉方式对表皮细胞增殖的影响。研究皮肤组织扩张与细胞机械牵拉对表皮细胞表达硫氧还蛋白(thioredoxin, TXN)的影响。质粒转染人永生化表皮细胞HaCat细胞使其过表达TXN,探讨TXN对细胞增殖的影响。一方面为研究人皮肤组织扩张术的分子机制提供新的线索；另一方面,将TXN作为新的潜在的分子靶点,为研究TXN的转基因治疗用于加快组织扩张速率提供新的实验证据。
方法
1、选取额部扩张器的患者,在附加切口分别取张力最大处扩张皮肤组织和邻近颞侧正常皮肤组织,分离出表皮层,利用Real Time PCR检测SMC5L1, PTH2_HUMAN, CKLF, TXN, S100B, TGIF2, SNAI2, PPP1R3C在mRNA水平的表达,利用免疫组织化学染色和Western Blot蛋白半定量检测TXN蛋白的表达。
2、根据国外报道常用的机械牵拉细胞培养模型的原理,结合皮肤组织扩张术的临床应用特点,自行设计并制作机械牵拉贴壁细胞培养模型；取植皮术中剩余的全厚皮,常规原代培养皮肤成纤维细胞和表皮细胞,取2-5代的细胞种植于模型的弹性膜上继续培养,观察成纤维细胞和表皮细胞在牵拉后细胞排列的差异。为研究不同牵拉方式对HaCat细胞增殖的影响,牵拉实验分为对照组(C,未牵拉组)、实验1组(M1,采用单次最大牵拉40%的延长度)、实验2组(M2,采用渐进式分四次达到最大牵拉40%的延长度,牵拉10%→停留→牵拉20%→停留→牵拉30%→停留→牵拉40%)。分别于牵拉后12h、24h、36h、48h、60h、72h时间点后,计数每个时间点的细胞数,利用MTT法检测细胞的增殖情况。于牵拉后48小时用Western blot蛋白半定量检测表皮细胞系在不同牵拉条件下和非牵拉条件下表达增殖细胞核抗原(proliferating cell nuclear antigen, PCNA)的情况。
3、采用自制的可调控细胞牵拉培养装置对HaCat细胞实行牵拉培养和无牵拉培养,Western blot蛋白半定量检测牵拉培养实施48小时后TXN蛋白表达情况。
4、用质粒pcDNA3.1-hTXN转染HaCat细胞,Western blot蛋白半定量检测转染48h和72h TXN和PCNA的表达。
结果
1、与前期基因芯片结果一致,定量PCR显示TXN mRNA相对表达水平在扩张皮肤组织和正常皮肤组织中分别为2.01±0.41和1.00±0.00,说明在mRNA水平扩张皮肤表皮层TXN表达较正常皮肤表皮层明显上调,有统计学意义(P<0.05)；TXN在表皮细胞、毛囊细胞和汗腺细胞的细胞核中高表达,扩张皮肤组织和正常皮肤组织中TXN阳性细胞率分别为(59.87±4.2)%和(41.52±8.4)%,扩张皮肤组织中TXN阳性细胞较正常皮肤组织高,有统计学意义(P<0.001)；扩张皮肤组织和正常皮肤表皮层中TXN蛋白表达量分别为1.48±0.24和0.65±0.14,显示扩张皮肤组织较正常皮肤组织表皮层显著升高,有统计学意义(P<0.05)。
2、研制两套牵拉细胞培养模型,一个是置入式牵拉细胞培养装置,该装置既可完成牵拉培养,又可置入普通培养皿,另一个是可控式牵拉细胞培养装置,可对细胞进行大幅度的持续性、渐进性的牵拉。二者方便消毒,易于观察细胞形态。
3、对人皮肤成纤维细胞和表皮细胞经牵拉培养发现,牵拉膜上的细胞生长状态良好,细胞沿牵拉方向呈明显极性排列,而未牵拉的皿内细胞排列杂乱无序。
4、与对照组(C组)相比,渐进牵拉组(M2组)和单次牵拉组(M1组)的细胞数目均增加,在48h处达到高峰；牵拉48h后,台盼蓝计数法结果显示三组的细胞数分别为(1.65±0.46)×105、(2.36±0.35)×105和(2.08±0.43)×105；MTT法结果显示三组的细胞吸光值分别为1.765±0.065、2.866±0.205和1.986±0.033。与C组相比,M2组细胞数增加明显,有统计学意义(P<0.05)。
5、比较不同牵拉方式作用48h后HaCat细胞表达PCNA蛋白的情况,正常组(C组)、渐进牵拉组(M2组)和单次牵拉组(M1组)中PCNA表达量分别为3.29±0.21、7.49±0.34和3.98±0.18,M2组与C组比较PCNA表达明显增加,有统计学意义(P<0.05)；M1组与C组比较PCNA表达有所增加,无统计学意义(P>0.05)。
6、比较机械应力牵拉培养48h后HaCat细胞表达TXN蛋白的情况,渐进式牵拉组(M组)较非牵拉组(C组)HaCat细胞表达TXN蛋白高,M组和C组中TXN表达量分别为0.79±0.09和0.14±0.05,M组与C组比较TXN表达明显增加,有统计学意义(P<0.05)。
7、将pcDNA3.1-hTXN质粒转染HaCat细胞,对照组、48h组和72h组TXN表达量分别为0.009±0.0003、0.048±0.0020和0.068±0.0110,对照组、48h组和72h组PCNA表达量分别为0.029±0.0001、0.076±0.0109和0.091±0.0093。72h组PCNA蛋白水平较C组表达增加明显,有统计学意义(P<0.05),48h组PCNA蛋白较C组有所增加,无统计学意义(P>0.05)。说明TXN的过表达会导致PCNA的表达增加。
结论
1、TXN在表皮细胞、毛囊细胞和汗腺细胞的细胞核中高表达,扩张皮肤组织表皮层中TXN mRNA和蛋白表达明显高于邻近正常皮肤组织表皮层；
2、可调控式牵拉培养装置可对贴壁细胞进行渐进式牵拉,渐进式牵拉方式更能充分模拟体内组织扩张术的过程。
3、机械应力可刺激表皮细胞表达TXN增多,TXN可能与机械应力促进细胞增殖有一定关系。
4、表皮细胞系中过表达TXN可能促进细胞的增殖,为后续研究组织扩张分子机制和转基因技术加快组织扩张速度提供实验基础。
Objective:
To establish a new cell culture model with mechanical stress that fully simulates the process of tissue expansion. To learn the influence of mechanical stress on human being epidermal keratinocyte proliferation in vitro by using different mechanical stretching methods. To investigate the influence of tissue expansion and mechanical stretching culture on the thioredoxin expression of skin keratinocyte. To explore effects of TXN overexpression on cell proliferation in HaCat cell. To provide new clues to studies on molecular mechanisms of tissue expansion and experimental evidence for clinic application of TXN gene therapy on speeding up the rate of tissue expansion by using TXN as a new potential molecular target.
Method:
1. Patients with frontal tissue expander were selected, from which skin tissues at the location of force-maximum and the temporalis normal skin tissues were taken at the extra-incision respectively to isolate epidermis. Utilize Real Time PCR to test the differential mRNA expression of SMC5L1, PTH2_HUMAN, CKLF, TXN, S100B, TGIF2, SNAI2, PPP1R3C. Test the protein expression of TXN by immunohischemistry and Western-Blot.
2. Based on mechanical stress cell culture theory regularly used in abroad reports, combined with the clinical characteristic of tissue expansion and skin stretching techniques, adherent cell culture model with mechanical stress was decised and constructed. Primary cultured skin fibroblasts and keratinocytes were obtained from full thickness skin of human being. The2-5passages fibroblasts and keratinocytes (cell density:5000-10000/cm2) digested with trypsin, were seeded on elastic membranes of the model. Then to investigate effects of different stretch methods to the keratinocyte on the cell proliferation, the cells were divided into three groups: group1、group2and control group. Group1was used as single maximum stress stretch (Ml group, only40%stretch), while group2was used as progressive and accumulative stress stretch way ((M2group.10%stretch→retention→20%stretch→retention→30%stretch→retention→40%stretch). The control group was no mechanical stress in the experiment. The number of HaCat cells was counted individually in12h、24h、36h、48h、60h、72h after the last of the mechanical stress, drawn out the proliferation curve and compared the cell proliferation between the tested group and the control one at different time with MTT. The total protein was also harvested from different groups in48h after the last mechanical stress. Cell cycle protein PCN A was analysised by Western Blot.
3. Stretched and unstretched HaCat cell culture were carried out by using self-made adjustable cell culture mechanical strain model. In48h after cultured in the stretching model, the total protein of HaCat cell was obtained and TXN protein was analysised by Western Blot.
4. The pcDNA3.1-hTXN recombinant vector was transfected into the HaCat cell to make the TXN enhanced.48h and72h after transfection, the protein levels of PCN A and TXN were analyzed by Western Blot.
Results:
1. In line with former data of microarray, the mRNA relative expression of TXN in epidermis of expanded skin tissue and normal skin tissue was2.01±0.4land1.00±0.00respectively. The mRNA of TXN in epidermis of expanded skin tissue increased compared with normal skin tissue group and there was statistical significance between groups(P<0.05). The TXN is mostly located in the nucleus of epidermal cells, hair follicles and sweat glands cell from the Immunohischemistry Staining. The positive cells rate of TXN expression in the expanded skin tissue and normal skin tissue was (59.87±4.2)%and (41.52±8.4)%respectively, and there was statistical significance between groups(P<0.001).The band density of TXN protein expression in epidermis of expanded skin tissue and normal skin tissue from Western Blot was1.48±0.24and0.65±0.14respectively, and there was statistical significance between groups(P<0.05).
2. Two sets of cell culture mechanical strain models were developed. One is the insertable cell culture mechanical strain model, which can be used to complete the stretch cultivation and at the same time can be insertable into the normal cell culture dish. The other is adjustable cell culture mechanical strain model, which can have substantial, continuous and gradual stretch on adherent cells. Both are easy to disinfect and are apt to observe cell morphology.
3. Results from the culture on human skin fibroblast cells and epidermal cells after stretches show that cells on the elastic membrane develop well.Cell morphology showed a polarity arranged along the stretch direction, while cells in the unstretched culture dishes arranged disorderly.
4. Compared with the control group (C), the number of cells in the accumulative stress stretch group (M2) and a single maximum stretch group (M1) were increased, peaked at48h After48h of stretching, the number of the cells in C group^M2group and M1group was (1.65±0.46)×105、(2.36±0.35)×105and (2.08±0.43)×105respectively from cell counting method, and1.765±0.065、2.866±0.205and1.986±0.033respectively from MTT method.Cell number of M2group apparently increased compared with C group, and there was statistical significance between groups(P<0.05).
5. After48h of stretching, the PCNA protein expression of C group、M2group and M1group was3.29±0.21、7.49±0.34and3.98±0.18. The PCNAprotein expression of M2group apparently increased compared with C group, and there was statistical significance between groups(P<0.05). The PCNA protein expression of M1group lightly increased compared with C group, and there was not statistical significance between groups(P>0.05).
6. After48h of stretching, the TXN protein expression of accumulative stress stretch group (M) and unstretched group(C) was0.79±0.09and0.14±0.05, and there was statistical significance between groups(P<0.05).
7. The TXN protein expression of untransfected group (C)、48h transfected group (48h) and72h transfected group (72h) was0.009±0.0003、0.048±0.0020and0.068±0.0110. The PCNA protein expression of untransfected group (C)、48h transfected group (48h) and72h transfected group (72h) was0.029±0.0001、0.076±0.0109and0.091±0.0093. The PCNA protein expression of72h group apparently increased compared with C group, and there was statistical significance between groups(P<0.05). The PCNA protein expression of48h group lightly increased compared with C group, and there was not statistical significance between groups(P>0.05).TXN overexpression will lead to increased expression of PCNA.
Conclusions:
1. TXN is overexpressed in the nucleus of epidermal cells, hair follicle cells and sweat gland cells. The TXN mRNA and protein expression in epidermis of expanded skin is significantly more than the neighbouring normal skin epidermis.
2. Adjustable cell culture mechanical strain model can help to have gradual traction to adherent cells, which can fully imitate the process of in vivo tissue expansion
3. Mechanical stress can stimulate TXN expression in skin keratinocytes, which suggests that there may exist some relevance between TXN and cell proliferation stimulated by mechanical stress.
4. Overexpression of TXN may stimulate cell proliferation of HaCat celL This study offers experimental basis for follow-up studies on the mechanism of cell proliferation with mechanical stress in the tissue expansion and the increase of tissue expansion speed by gene therapy.
建立充分模拟皮肤组织扩张过程的贴壁细胞机械牵拉培养体外模型；研究不同牵拉方式对表皮细胞增殖的影响。研究皮肤组织扩张与细胞机械牵拉对表皮细胞表达硫氧还蛋白(thioredoxin, TXN)的影响。质粒转染人永生化表皮细胞HaCat细胞使其过表达TXN,探讨TXN对细胞增殖的影响。一方面为研究人皮肤组织扩张术的分子机制提供新的线索；另一方面,将TXN作为新的潜在的分子靶点,为研究TXN的转基因治疗用于加快组织扩张速率提供新的实验证据。
方法
1、选取额部扩张器的患者,在附加切口分别取张力最大处扩张皮肤组织和邻近颞侧正常皮肤组织,分离出表皮层,利用Real Time PCR检测SMC5L1, PTH2_HUMAN, CKLF, TXN, S100B, TGIF2, SNAI2, PPP1R3C在mRNA水平的表达,利用免疫组织化学染色和Western Blot蛋白半定量检测TXN蛋白的表达。
2、根据国外报道常用的机械牵拉细胞培养模型的原理,结合皮肤组织扩张术的临床应用特点,自行设计并制作机械牵拉贴壁细胞培养模型；取植皮术中剩余的全厚皮,常规原代培养皮肤成纤维细胞和表皮细胞,取2-5代的细胞种植于模型的弹性膜上继续培养,观察成纤维细胞和表皮细胞在牵拉后细胞排列的差异。为研究不同牵拉方式对HaCat细胞增殖的影响,牵拉实验分为对照组(C,未牵拉组)、实验1组(M1,采用单次最大牵拉40%的延长度)、实验2组(M2,采用渐进式分四次达到最大牵拉40%的延长度,牵拉10%→停留→牵拉20%→停留→牵拉30%→停留→牵拉40%)。分别于牵拉后12h、24h、36h、48h、60h、72h时间点后,计数每个时间点的细胞数,利用MTT法检测细胞的增殖情况。于牵拉后48小时用Western blot蛋白半定量检测表皮细胞系在不同牵拉条件下和非牵拉条件下表达增殖细胞核抗原(proliferating cell nuclear antigen, PCNA)的情况。
3、采用自制的可调控细胞牵拉培养装置对HaCat细胞实行牵拉培养和无牵拉培养,Western blot蛋白半定量检测牵拉培养实施48小时后TXN蛋白表达情况。
4、用质粒pcDNA3.1-hTXN转染HaCat细胞,Western blot蛋白半定量检测转染48h和72h TXN和PCNA的表达。
结果
1、与前期基因芯片结果一致,定量PCR显示TXN mRNA相对表达水平在扩张皮肤组织和正常皮肤组织中分别为2.01±0.41和1.00±0.00,说明在mRNA水平扩张皮肤表皮层TXN表达较正常皮肤表皮层明显上调,有统计学意义(P<0.05)；TXN在表皮细胞、毛囊细胞和汗腺细胞的细胞核中高表达,扩张皮肤组织和正常皮肤组织中TXN阳性细胞率分别为(59.87±4.2)%和(41.52±8.4)%,扩张皮肤组织中TXN阳性细胞较正常皮肤组织高,有统计学意义(P<0.001)；扩张皮肤组织和正常皮肤表皮层中TXN蛋白表达量分别为1.48±0.24和0.65±0.14,显示扩张皮肤组织较正常皮肤组织表皮层显著升高,有统计学意义(P<0.05)。
2、研制两套牵拉细胞培养模型,一个是置入式牵拉细胞培养装置,该装置既可完成牵拉培养,又可置入普通培养皿,另一个是可控式牵拉细胞培养装置,可对细胞进行大幅度的持续性、渐进性的牵拉。二者方便消毒,易于观察细胞形态。
3、对人皮肤成纤维细胞和表皮细胞经牵拉培养发现,牵拉膜上的细胞生长状态良好,细胞沿牵拉方向呈明显极性排列,而未牵拉的皿内细胞排列杂乱无序。
4、与对照组(C组)相比,渐进牵拉组(M2组)和单次牵拉组(M1组)的细胞数目均增加,在48h处达到高峰；牵拉48h后,台盼蓝计数法结果显示三组的细胞数分别为(1.65±0.46)×105、(2.36±0.35)×105和(2.08±0.43)×105；MTT法结果显示三组的细胞吸光值分别为1.765±0.065、2.866±0.205和1.986±0.033。与C组相比,M2组细胞数增加明显,有统计学意义(P<0.05)。
5、比较不同牵拉方式作用48h后HaCat细胞表达PCNA蛋白的情况,正常组(C组)、渐进牵拉组(M2组)和单次牵拉组(M1组)中PCNA表达量分别为3.29±0.21、7.49±0.34和3.98±0.18,M2组与C组比较PCNA表达明显增加,有统计学意义(P<0.05)；M1组与C组比较PCNA表达有所增加,无统计学意义(P>0.05)。
6、比较机械应力牵拉培养48h后HaCat细胞表达TXN蛋白的情况,渐进式牵拉组(M组)较非牵拉组(C组)HaCat细胞表达TXN蛋白高,M组和C组中TXN表达量分别为0.79±0.09和0.14±0.05,M组与C组比较TXN表达明显增加,有统计学意义(P<0.05)。
7、将pcDNA3.1-hTXN质粒转染HaCat细胞,对照组、48h组和72h组TXN表达量分别为0.009±0.0003、0.048±0.0020和0.068±0.0110,对照组、48h组和72h组PCNA表达量分别为0.029±0.0001、0.076±0.0109和0.091±0.0093。72h组PCNA蛋白水平较C组表达增加明显,有统计学意义(P<0.05),48h组PCNA蛋白较C组有所增加,无统计学意义(P>0.05)。说明TXN的过表达会导致PCNA的表达增加。
结论
1、TXN在表皮细胞、毛囊细胞和汗腺细胞的细胞核中高表达,扩张皮肤组织表皮层中TXN mRNA和蛋白表达明显高于邻近正常皮肤组织表皮层；
2、可调控式牵拉培养装置可对贴壁细胞进行渐进式牵拉,渐进式牵拉方式更能充分模拟体内组织扩张术的过程。
3、机械应力可刺激表皮细胞表达TXN增多,TXN可能与机械应力促进细胞增殖有一定关系。
4、表皮细胞系中过表达TXN可能促进细胞的增殖,为后续研究组织扩张分子机制和转基因技术加快组织扩张速度提供实验基础。
Objective:
To establish a new cell culture model with mechanical stress that fully simulates the process of tissue expansion. To learn the influence of mechanical stress on human being epidermal keratinocyte proliferation in vitro by using different mechanical stretching methods. To investigate the influence of tissue expansion and mechanical stretching culture on the thioredoxin expression of skin keratinocyte. To explore effects of TXN overexpression on cell proliferation in HaCat cell. To provide new clues to studies on molecular mechanisms of tissue expansion and experimental evidence for clinic application of TXN gene therapy on speeding up the rate of tissue expansion by using TXN as a new potential molecular target.
Method:
1. Patients with frontal tissue expander were selected, from which skin tissues at the location of force-maximum and the temporalis normal skin tissues were taken at the extra-incision respectively to isolate epidermis. Utilize Real Time PCR to test the differential mRNA expression of SMC5L1, PTH2_HUMAN, CKLF, TXN, S100B, TGIF2, SNAI2, PPP1R3C. Test the protein expression of TXN by immunohischemistry and Western-Blot.
2. Based on mechanical stress cell culture theory regularly used in abroad reports, combined with the clinical characteristic of tissue expansion and skin stretching techniques, adherent cell culture model with mechanical stress was decised and constructed. Primary cultured skin fibroblasts and keratinocytes were obtained from full thickness skin of human being. The2-5passages fibroblasts and keratinocytes (cell density:5000-10000/cm2) digested with trypsin, were seeded on elastic membranes of the model. Then to investigate effects of different stretch methods to the keratinocyte on the cell proliferation, the cells were divided into three groups: group1、group2and control group. Group1was used as single maximum stress stretch (Ml group, only40%stretch), while group2was used as progressive and accumulative stress stretch way ((M2group.10%stretch→retention→20%stretch→retention→30%stretch→retention→40%stretch). The control group was no mechanical stress in the experiment. The number of HaCat cells was counted individually in12h、24h、36h、48h、60h、72h after the last of the mechanical stress, drawn out the proliferation curve and compared the cell proliferation between the tested group and the control one at different time with MTT. The total protein was also harvested from different groups in48h after the last mechanical stress. Cell cycle protein PCN A was analysised by Western Blot.
3. Stretched and unstretched HaCat cell culture were carried out by using self-made adjustable cell culture mechanical strain model. In48h after cultured in the stretching model, the total protein of HaCat cell was obtained and TXN protein was analysised by Western Blot.
4. The pcDNA3.1-hTXN recombinant vector was transfected into the HaCat cell to make the TXN enhanced.48h and72h after transfection, the protein levels of PCN A and TXN were analyzed by Western Blot.
Results:
1. In line with former data of microarray, the mRNA relative expression of TXN in epidermis of expanded skin tissue and normal skin tissue was2.01±0.4land1.00±0.00respectively. The mRNA of TXN in epidermis of expanded skin tissue increased compared with normal skin tissue group and there was statistical significance between groups(P<0.05). The TXN is mostly located in the nucleus of epidermal cells, hair follicles and sweat glands cell from the Immunohischemistry Staining. The positive cells rate of TXN expression in the expanded skin tissue and normal skin tissue was (59.87±4.2)%and (41.52±8.4)%respectively, and there was statistical significance between groups(P<0.001).The band density of TXN protein expression in epidermis of expanded skin tissue and normal skin tissue from Western Blot was1.48±0.24and0.65±0.14respectively, and there was statistical significance between groups(P<0.05).
2. Two sets of cell culture mechanical strain models were developed. One is the insertable cell culture mechanical strain model, which can be used to complete the stretch cultivation and at the same time can be insertable into the normal cell culture dish. The other is adjustable cell culture mechanical strain model, which can have substantial, continuous and gradual stretch on adherent cells. Both are easy to disinfect and are apt to observe cell morphology.
3. Results from the culture on human skin fibroblast cells and epidermal cells after stretches show that cells on the elastic membrane develop well.Cell morphology showed a polarity arranged along the stretch direction, while cells in the unstretched culture dishes arranged disorderly.
4. Compared with the control group (C), the number of cells in the accumulative stress stretch group (M2) and a single maximum stretch group (M1) were increased, peaked at48h After48h of stretching, the number of the cells in C group^M2group and M1group was (1.65±0.46)×105、(2.36±0.35)×105and (2.08±0.43)×105respectively from cell counting method, and1.765±0.065、2.866±0.205and1.986±0.033respectively from MTT method.Cell number of M2group apparently increased compared with C group, and there was statistical significance between groups(P<0.05).
5. After48h of stretching, the PCNA protein expression of C group、M2group and M1group was3.29±0.21、7.49±0.34and3.98±0.18. The PCNAprotein expression of M2group apparently increased compared with C group, and there was statistical significance between groups(P<0.05). The PCNA protein expression of M1group lightly increased compared with C group, and there was not statistical significance between groups(P>0.05).
6. After48h of stretching, the TXN protein expression of accumulative stress stretch group (M) and unstretched group(C) was0.79±0.09and0.14±0.05, and there was statistical significance between groups(P<0.05).
7. The TXN protein expression of untransfected group (C)、48h transfected group (48h) and72h transfected group (72h) was0.009±0.0003、0.048±0.0020and0.068±0.0110. The PCNA protein expression of untransfected group (C)、48h transfected group (48h) and72h transfected group (72h) was0.029±0.0001、0.076±0.0109and0.091±0.0093. The PCNA protein expression of72h group apparently increased compared with C group, and there was statistical significance between groups(P<0.05). The PCNA protein expression of48h group lightly increased compared with C group, and there was not statistical significance between groups(P>0.05).TXN overexpression will lead to increased expression of PCNA.
Conclusions:
1. TXN is overexpressed in the nucleus of epidermal cells, hair follicle cells and sweat gland cells. The TXN mRNA and protein expression in epidermis of expanded skin is significantly more than the neighbouring normal skin epidermis.
2. Adjustable cell culture mechanical strain model can help to have gradual traction to adherent cells, which can fully imitate the process of in vivo tissue expansion
3. Mechanical stress can stimulate TXN expression in skin keratinocytes, which suggests that there may exist some relevance between TXN and cell proliferation stimulated by mechanical stress.
4. Overexpression of TXN may stimulate cell proliferation of HaCat celL This study offers experimental basis for follow-up studies on the mechanism of cell proliferation with mechanical stress in the tissue expansion and the increase of tissue expansion speed by gene therapy.
引文
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