[Study on bone marrow mesenchymal stem cells transfected with adenovirus hepatocyte growth factor gene promoting wounds repair in diabetic rats].
OBJECTIVE
To explore the effects of bone marrow mesenchymal stem cells (BMSCs) transfected with adenovirus hepatocyte growth factor (Ad-HGF) on wound repair in diabetic rats. MRTHODS: BMSCs from male Wistar rats were isolated by density gradient centrifugation, cultured, and transfected with Ad-HGF. The multiplicity of infection was 100. Diabetic models were established in 20 female Wistar rats by diets in high fat and sugar plus intraperitoneal injection of streptozotocin (30 mg/kg). Then 2 full-thickness skin wounds (approximately 1.5 cm in diameter) were made on the dorsum. The rats were randomly divided into 4 groups (n = 5 rats). After wounding, the 0.3 mL suspensions of BMSCs (group A), Ad-HGF (group B), BMSCs transfected with Ad-HGF (group C), and PBS (group D) were injected directly into the dermal of wounds. The transverse diameter and longitudinal diameter of wound were measured at 21 days after treatment. At 7 days and 28 days after treatment, HE staining was performed to evaluate wound healing. The contents of hydroxyproline and advanced glycosylation end products (AGEs) in the wounds were measured by enzyme linked immunosorbent assay and fluorospectrophotometer, respectively, at 3, 7, 14, and 28 days after treatment.


RESULTS
At 21 days after treatment, the wounds almost healed in group C, and the transverse diameter and longitudinal diameter were 0 and (0.110 +/- 0.024) cm, respectively. But the wounds healed partially in groups A, B, and D, and the transverse diameter and longitudinal diameter were (0.470 +/- 0.051) cm and (0.590 +/- 0.041) cm, (0.390 +/- 0.042) cm and (0.480 +/- 0.032) cm, and (0.700 +/- 0.068) cm and (0.820 +/- 0.068) cm, respectively. There were significant differences in wound healing between group C and groups A, B, and D (P < 0.05). The wound healing time of group C [(20.5 +/- 1.9) days] was significantly shorter (P < 0.05) than those of groups A, B, and D [(28.3 +/- 1.9), (25.9 +/- 2.3), and (36.6 +/- 5.1) days]. At 7 days, the HE staining showed that evident epidermis transportation, collagen formation, and leukocytes infiltration were observed in group C. At 28 days, the HE staining showed that the epidermis in group C was significantly thinner and more regular than those in other groups, and the decreased collagen and many small vessels were observed in group C. The content of hydroxyproline in group C was higher than those in groups A, B, and D at 7 days and 14 days (P < 0.05). The contents of AGEs in group C was lower than those in groups A, B, and D at 14 days and 28 days (P < 0.05). CONCLUSION Transplantation of BMSCs transfected with Ad-HGF can accelerate the wounds repair in diabetic rats.
Experimental Center of Medicine, Lanzhou Military General Hospital of Chinese PLA, Key Laboratory of Stem Cell and Gene Medicine of Gansu Province, Lanzhou Gansu 730050, PR China.
Xiaoqin
X
Ha
Qiang
Q
Yin
Fang
F
Dong
Qinghua
Q
Jia
Tongde
T
Lv
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002478:Cells, Cultured; D003921:Diabetes Mellitus, Experimental; D005260:Female; D017228:Hepatocyte Growth Factor / Q000235:genetics / Q000494:pharmacology*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051381:Rats; D017208:Rats, Wistar; D014162:Transfection; D014945:Wound Healing
D017228:Hepatocyte Growth Factor