Objective To investigate the effects of the recombinant plasmid pIRES-hBMP-2-hVEGF165 on differentiation and maturation of hBMSCs in vitro. Methods The co-expressing vector of hBMP-2 and hVEGF165 was constructed. The BMSCs were isolated and cultured from health adult human denoted marrow. By the l ipofection method, the reconstructed plasmids pIRES-hBMP-2-hVEGF165, pIRES-hBMP-2, pIRES-hVEGF165 and pIRES neo empty vector, weretransfected to hBMSCs (groups A, B, C and D). The untransfected cells were harvested as control group (group E). After4 weeks of culture, RT-PCR was employed to assay the hBMP-2, hVEGF165 and osteocalcin mRNA expression in hBMSCs. The expressions of hBMP-2 and hVEGF165 of BMSCs were assayed by Western blot. The level of ALP activities of BMSCs was determined. Col I was also determined by immunohistochemical staining. Results Compared to group E, the hBMSCs in group A secreted high level of hBMP-2, hVEGF165, Col I and osteocalcin; osteocalcin and Col I expressed at high level in group B, and hVEGF165 expressed at high level in group C. Otherwise, the expression of hVEGF165 in group B and the expressions of hBMP-2 and Col I in group C resemble to that of groups D and E, no expression or few expression was observed. The activities of ALP in groups A, B, C, D and E were 0.91 ± 0.03, 0.90 ± 0.02, 0.64 ± 0.03, 0.67 ± 0.01 and 0.66 ± 0.02, respectively. The activity of ALP of groups A and B were significantly increased compared with that of group E (P lt; 0.05); there was no significant difference among groups C, D and E (P gt; 0.05). Conclusion The recombinant plasmid pIRES-hBMP-2-hVEGF165 can be successfully transfected into BMSCs with cation l iposome-mediated transfection method, the exogenous hBMP-2 and hVEGF165 genes can be expressed constitutively in the transfected BMSCs, and it can enhance the differentiation abil ities of BMSCs.
Objective To explore the therapeutic effect of recombinant human epidermal growth factor (rhEGF) for burn wounds of degree II in the elderly patients. Methods From February 2003 to October 2008, 80 patientes with burn wounds of degree II were treated and randomly divided into two groups (n=40). In treatment group, there were 24 males and 16 females with an average age of 70 years (60-86 years), including 20 cases of superficial II degree and 20 cases of deep II degree.Burn wounds were caused by flame in 23 cases, by hot l iquid in 16 cases, and by electricity in 1 case. The mean time from injury to hospital ization was (2.87 ± 2.57) hours. The wounds were treated with silver sulfadiazine (SD-Ag) and rhEGF. In control group, there were 18 males and 22 females with an average age of 69 years (61-83 years), including 19 cases of superficial II degree and 21 cases of deep II degree. Burn wounds were caused by flame in 23 cases, by hot l iquid in 14 cases, by electricity in 2 cases, and by chemistry in 1 case. The mean time from injury to hospital ization was (3.39 ± 3.33) hours. The wounds were treated with SD-Ag. The dressing was changed every day until wounds heal ing. There were no significant differences in general data between two groups (P gt; 0.05). Results Wound did not heal in 1 case (deep II degree) of treatment group and in 5 cases (deep II degree) of control group over 40 days and free skin graft was used to repair wound. One case (superficial II degree ) in control group gave up treatment. One case (deep II degree) died of pulmonary infection in treatment group. These cases were excluded and 72 cases were analysed. No other side reactions were observed in teatment group except for flash stabbing pain (4 cases) and pruritus (2 cases). Wound infection occurred in 5 cases of the control group and in 3 cases of the treatment group, and wound healed after symptomatic treatment. The heal ing time of burn wound was (14.30 ± 1.26) days (superficial II degree) and (26.11 ± 2.97) days (deep II degree) in the treatment group, was (16.22 ± 1.40) days (superficial II degree) and (29.13 ± 4.99) days (deep II degree) in control group, showing significant difference between two groups (P lt; 0.05). Conclusion Incombined treatment, rhEGF can promote the heal ing of burn wounds of degree II in the elderly patients.
Objective To observe the effect of continuous elastic outside distraction on the change of collagen content in female mini pig’s ni pples and their supporting tissues, and to investigate the mechanism of continuous elastic outside distraction correcting inverted ni pples. Methods Three 3-month-old female mini pigs (weighing 18.5-22.0 kg), which had 12 nipples, were employed. Four nipples of each minipig were not treated as control group (n=12), and the other nipples were continuously distracted with inverted nipple correction instruments as experimental group (n=24). The nipple specimens were harvested at 2, 4, 8, and 12 weeks after distraction and HE staining was performed to observe the change oftheir tissue structure. And saturated picric acid sirius red staining was used to observe the distribution and content of collagen types I and III, image analysis software for quantitative analysis. Results The control group had normal structure of epidermis at all time points. In experimental group, the epidermis thickened; basal cells, fibroblasts, and capillary significantly prol iferated along with the times; and the content and the density of collagen types I and III increased gradually. There were significant differences in collagen type I at 4, 8, and 12 weeks, and in collagen type III at 2, 4, 8, and 12 weeks between 2 groups (P lt; 0.01). There were significant differences in the ratio of collagen type I to III at 2 and 4 weeks between 2 groups (P lt; 0.05). Conclusion Continuous elastic outside distraction can increase the quantity of collagen types I and III in the tissue, the thickness of the dermis, and the height of the nipple, which may be one of key mechanisms of correction the inverted nipple by continuous elastic outside distraction.
Objective To investigate the effectiveness of free anterolateral thigh flap (ALTF) with fascia lata in repairing diabetic foot ulcers (DFUs) with bone exposure. Methods Between January 2019 and January 2021, 20 patients with DFUs with bone exposure were admitted. There were 17 males and 3 females with a median age of 57.5 years (range, 48-76 years). There were 10 cases of Wagner grade 3 and 10 cases of grade 4. The DFUs formed 1 to 14 months, with a median time of 3 months. The patients underwent CT angiography, which showed extensive atherosclerosis in both lower limbs; 6 of them were severely narrowed or occluded and underwent percutaneous transluminal angioplasty. The size of wound ranged from 7 cm×6 cm to 27 cm×10 cm after applied first-stage debridement combined with vacuum sealing drainage treatment. In the second-stage, free ALTF with fascia lata was used to repair wounds and partial defects of tendons. The size of flap ranged from 8 cm×5 cm to 28 cm×11 cm. The wound of the donor site was sutured directly. The survival of the flap, the healing time of the wound, and the complications were recorded. The laser speckle blood flow imaging system was used to detect the blood perfusion of the flap and the skin around the flap at 2 weeks and 6 months after operation. The foot function was evaluated by American Orthopaedic Foot and Ankle Society (AOFAS) score at 6 months after operation. Results After operation, effusion under the flap happened in 6 cases, which cured after symptomatic treatment. Flaps survived completely in 14 cases. The tissue necrosis at the edges of the flaps occurred in 3 cases and healed after dressing changes. Venous crisis of flaps occurred in 3 cases, of which 1 case was completely necrotic after exploration, and the other 2 cases were partially alive. The wounds of 3 cases were repaired with skin grafts after debridement and dressing. The flap survival rate was 95.0%, and the limb salvage rate was 100%. The wound healing time after flap transplantation was 14-30 days, with an average of 19.1 days. Two patients had recurrence of peripheral skin ulcers of the flaps within 1 month after healing, which healed after conservative dressing changes. Eighteen cases of incisions at donor site healed by first intention, 2 cases had local skin necrosis and healed by debridement and suture. All patients were followed up 6-30 months, with a median time of 11 months. The texture, appearance, and elasticity of the flaps were good. All patients could walk alone without pain. At 6 months after operation, the AOFAS score was 75.9±11.9, which was significantly different from that (44.7±18.4) before operation (t=−7.025, P=0.000). The blood perfusion value increased from (38.1±7.8) PU at 2 weeks to (42.7±10.3) PU, and the difference was significant (t=−4.680, P=0.001). Conclusion Free ALTF with fascia lata has a rich blood supply and a high survival rate. It can be used to repair DFUs with bone exposure. After the free skin flap healed, it can promote revascularization of the affected foot, reduce the probability of ulcer recurrence, and avoid amputation.