脂肪间充质干细胞对巴马小型猪自体皮肤移植愈合过程的影响

doi:10.11843/j.issn.0366-6964.2024.07.037

摘要/Abstract

摘要：

拟应用脂肪间充质干细胞(adipose derived mesenchymal stem cells，ADSCs)干预小型猪自体皮肤移植实验动物模型，观察其对自体皮肤移植愈合过程中的作用效果。对9头小型猪建立自体皮肤移植动物模型，将小型猪随机分为3组：模型组(Model)、磷酸盐缓冲液组(PBS)、脂肪间充质干细胞组(ADSCs)。各组分别于术前，术后7、14、21和28 d，采集血液和皮肤组织样本，并观察、拍照、记录移植皮片和周围组织临床变化情况。通过对移植皮片表观检查、病理组织学检测，血常规以及血清生化检测，移植皮片组织氧化应激、炎症、再生相关指标检测及血流恢复情况，综合评价ADSCs对小型猪自体皮肤移植创口愈合的影响。临床观察发现，术后ADSCs组皮片恢复速度优于其他两组，在术后28 d，ADSCs组基本恢复正常皮肤状态。在氧化应激方面，ADSCs组SOD、GSH含量显著升高(0.01＜P < 0.05)，MDA显著下降(0.01＜P < 0.05)。在炎症方面，ADSCs组显著降低TNF-α、IL-1β含量，IL-10在术后7、14 d极显著升高(P＜0.01)。在皮肤组织再生方面，ADSCs组VEGF、bFGF、TGFβ3和CD31的表达量显著上升(0.01＜P＜0.05)。在血流恢复方面，ADSCs在愈合过程中血流灌注百分比显著高于其他两组(0.01＜P < 0.05)。综上表明，在小型猪自体皮肤移植动物模型中，ADSCs能够加速皮肤组织病理损伤的恢复，降低氧化应激反应及炎症反应，促进皮肤组织再生，血管新生。表明ADSCs具有抗氧化、抗炎和促进组织再生的功能对小型猪自体皮肤移植具有良好的治疗效果，本试验为干细胞在自体皮肤移植和异体皮肤移植的临床应用提供了实验支撑和科学依据。

关键词: 小型猪, 脂肪间充质干细胞, 自体皮肤移植, 创伤愈合

Abstract:

This study aims to use adipose derived mesenchymal stem cells (ADSCs) to intervene in an experimental animal model of autologous skin transplantation in small pigs, and observe their effects on the healing process of autologous skin transplantation. The animal model of autologous skin transplantation of 9 miniature pigs was established, and the miniature pigs were randomly divided into 3 groups: model group (Model), phosphate buffer group (PBS), and adipose mesenchymal stem cell group (ADSCs). Blood and skin tissue samples were collected before surgery and 7, 14, 21, and 28 days after surgery, respectively, and clinical changes of grafts and surrounding tissues were observed, photographed, and recorded. The effect of ADSCs on wound healing of mini pig autologous skin transplantation was comprehensively evaluated through skin graft apparent examination, histopathological examination, blood routine examination and serum biochemical examination, skin graft tissue oxidative stress, inflammation, regeneration and blood flow recovery. The researchers observed that the skin recovery speed of ADSCs group was better than that of the other two groups, and the ADSCs group basically returned to normal skin status 28 days after surgery. In terms of oxidative stress, SOD and GSH contents in ADSCs group were significantly increased (0.01 < P < 0.05), while MDA contents were significantly decreased (0.01 < P < 0.05). In terms of inflammation, the contents of TNF-α and IL-1β in ADSCs group were significantly decreased, and IL-10 was significantly increased on 7 and 14 days after surgery (P < 0.01). In terms of skin tissue regeneration, the expression levels of VEGF, bFGF, TGFβ3 and CD31 in ADSCs group were significantly increased (0.01 < P < 0.05). In terms of blood flow recovery, the percentage of blood perfusion of ADSCs during healing was significantly higher than that of the other two groups (0.01 < P < 0.05). To sum up, ADSCs can accelerate the recovery of skin tissue pathological damage, reduce oxidative stress reaction promote skin tissue regeneration and Angiogenesis in the miniature pig autologous skin transplantation animal model. The results indicated that ADSCs had the functions of anti-oxidation, anti-inflammation and promoting tissue regeneration, and had good therapeutic effect on autologous skin transplantation of miniature pigs. This experiment provided experimental support and scientific basis for the clinical application of stem cells in autologous skin transplantation and allogeneic skin transplantation.

Key words: small pigs, adipose mesenchymal stem cells, autologous skin transplantation, wound healing

中图分类号:

S857.132

吕英光, 焦广明, 桑金芳, 寇志鹏, 刘涛, 王月, 陆翔宇, 朴晨曦, 马亚军, 张建涛, 王洪斌. 脂肪间充质干细胞对巴马小型猪自体皮肤移植愈合过程的影响[J]. 畜牧兽医学报, 2024, 55(7): 3193-3204.

Yingguang LÜ, Guangming JIAO, Jinfang SANG, Zhipeng KOU, Tao LIU, Yue WANG, Xiangyu LU, Chenxi PIAO, Yajun MA, Jiantao ZHANG, Hongbin WANG. The Effect of Adipose Mesenchymal Stem Cells on the Healing Process of Autologous Skin Transplantation in Bama Miniature Pigs[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(7): 3193-3204.

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指标 Index	组别 Group	术前 Before operation	术后7 d 7 d after surgery	术后14 d 14 d after surgery	术后21 d 21 d after surgery	术后28 d 28 d after surgery
白细胞 WBC	Model	11.57±3.77	17.75±6.21	14.61±3.64	12.37±3.70	11.83±3.22
	PBS	12.18±3.90	19.53±6.57	19.13±3.27	15.89±3.51	13.75±3.36
	ADSCs	14.53±2.74	22.17±3.32	16.91±2.87	16.53±2.69	12.96±2.77
中性粒细胞 NE	Model	5.47±0.64	9.91±2.73	6.87±1.96	6.36±1.24	5.53±0.88
	PBS	6.35±1.16	12.75±3.41	12.85±1.69	9.65±1.51	7.55±1.03
	ADSCs	7.56±0.85	14.01±3.02	9.24±1.73	9.84±1.26	7.62±1.44
淋巴细胞 LY	Model	5.15±2.49	6.62±3.65	5.37±2.77	5.02±2.33	4.89±2.41
	PBS	4.72±2.36	5.05±3.58	4.65±2.96	4.41±2.12	4.95±1.87
	ADSCs	4.33±0.82	7.44±1.53	6.12±1.20	4.46±0.92	4.13±0.95

指标 Index	组别 Group	术前 Before operation	术后7 d 7 d after surgery	术后14 d 14 d after surgery	术后21 d 21 d after surgery	术后28 d 28 d after surgery
总蛋白 TP	Model	81.90±2.81	73.10±5.44	71.80±3.73	71.30±3.72	71.60±3.17
	PBS	75.50±3.85	71.30±6.51	65.90±3.31	71.50±3.49	65.80±3.34
	ADSCs	80.20±2.63	68.20±3.26	74.10±2.80	69.50±2.71	64.90±2.67
白蛋白 ALB	Model	34.90±1.31	32.90±2.15	32.90±1.83	34.00±1.07	34.70±1.06
	PBS	38.00±2.47	34.70±3.63	35.50±2.15	34.60±1.16	35.60±1.58
	ADSCs	40.10±1.35	34.10±1.71	36.20±1.60	35.40±1.23	33.00±1.33
球蛋白 GLO	Model	47.00±1.32	40.20±3.07	38.90±1.77	37.30±2.33	36.90±2.01
	PBS	37.50±1.26	36.60±2.47	30.40±1.16	36.90±2.12	30.20±1.67
	ADSCs	40.10±1.04	34.10±1.37	37.90±1.39	34.10±0.97	31.90±1.16

时间Time	模型组Model group	PBS组PBS group	ADSCs组ADSCs group
术前Before operation	21.82±1.84	22.42±3.09	21.07±2.88
7 d	7.70±1.21	8.35±1.92	10.62±1.65
14 d	11.36±2.27	12.06±3.13	13.54±1.81
21 d	14.96±0.83	16.25±2.10	19.87±2.18
28 d	19.18±0.83	21.11±2.34	20.51±2.25

时间/d Time	模型组Model group	PBS组PBS group	ADSCs组ADSCs group
7	35.15±3.05	36.92±3.36	51.48±13.92
14	51.77±6.17	53.27±6.65	65.69±17.10
21	68.84±6.17	72.52±1.67	94.57±2.80^##^**
28	88.08±3.74	94.42±3.81	97.59±2.87^*

评分Score	皮肤完整程度Skin integrity
0	完整的移植物
1	第一次出现排斥反应且排斥面积≤25%
2	排斥面积>25%且≤50%
3	排斥面积>50%且≤75%
4	排斥面积>75%且≤95%
5	排斥面积>95%