Autologous fat transplantation for the treatment of abdominal wall scar adhesions after cesarean section.

Cesarean section results in scarring, which usually leads to adhesion between the subcutaneous fat and the abdominal wall muscle. The present study aimed to evaluate the therapeutic effect of autologous fat grafting on scar adhesion to the abdominal wall after cesarean section. Thirty-six patients with scar adhesion to the abdominal wall after cesarean section were recruited and treated between October 2013 and December 2015. The adhesion between the subcutaneous fat and the abdominal wall muscle was carefully separated through a small incision in the original scar to form multiple subcutaneous tunnels. Aspirated fat was injected into the scar lesion and subcutaneous tunnels, and the wound was then sutured. The clinical outcome was evaluated by comparing the pretreatment and 1-year posttreatment photographs and Patient and Observer Scar Assessment Scale (POSAS) scores. All patients had a marked improvement in the appearance, texture, and depression of the scar during 12 months of follow-up. The 1-year posttreatment POSAS scores for the color, pain, pruritus, hardness, fullness, mobility, and appearance of the scar were significantly decreased compared with the pretreatment scores. Hematoxylin-eosin staining revealed adipocyte-like cells in treated scar tissue specimens obtained 1 year after treatment. None of the patients reported severe adverse reactions. Autologous fat grafting combined with adhesion release may be a good treatment option for abdominal wall scarring after cesarean section. This method is minimally invasive and effective in achieving good functional and esthetic outcomes.

Allogeneic Platelet-Rich Plasma Therapy as an Effective and Safe Adjuvant Method for Chronic Wounds.

BACKGROUND: Platelet-rich plasma (PRP) improves the healing of refractory wounds, and its application is receiving more attention in the field of wound repair. However, when a patient's condition is very poor, it may be difficult to provide whole blood to harvest autologous PRP. METHODS: We evaluated the efficacy and safety of allogeneic PRP in the field of chronic refractory wound repair. Sixty patients (39 males and 21 females, 57 ± 10 y old) with chronic wounds were enrolled in this prospective, randomized, single-center study during January 2014 to January 2018. Their wounds were treated by standard care. The patients with chronic refractory wounds were divided into allogeneic PRP treatment and control groups on the basis of the presence or absence of allogeneic PRP in wounds after debridement, respectively. Allogeneic PRP was prepared by collecting whole blood from healthy individuals and two-step centrifugation. Clinical effects were evaluated by visually observing wound conditions and objectively assessing wound surfaces. RESULTS: After 30 d of treatment, the allogeneic PRP-treated group showed bright red granulation that bled easily with reduced inflammatory exudation. No rejection reactions were observed. The rate of chronic wound healing was much faster in the allogeneic PRP-treated group than that in the control group. CONCLUSIONS: The present study shows that combined treatment of chronic wounds by standard care and allogeneic PRP significantly shortens healing time, suggesting that allogeneic PRP is an effective, safe adjuvant treatment for chronic wounds.

Antiaging Properties of Exosomes from Adipose-Derived Mesenchymal Stem Cells in Photoaged Rat Skin.

Adipose-derived stem cells (ADSCs) have been documented as possible candidates for skin rejuvenation. However, the effects of ADSC-derived exosomes on photoaged skin remain to be fully elucidated. This study was aimed at determining the antiaging effects of ADSC-derived exosomes on photoaged skin. Human ADSCs were isolated from the adipose tissue of healthy women and cultured in vitro. Then, exosomes were extracted from the cultured ADSCs, purified by ultracentrifugation, and verified by examination of cell morphology using transmission electron microscopy and the identification of specific biomarkers. Meanwhile, the optimal exosome concentration and treatment time were selected. The photoaged skin model was created by subjecting Sprague-Dawley rats to ultraviolet B radiation. Exosomes were injected into the photoaged skin in a single therapeutic dose. The thickness of the epidermis and dermis was observed by HE staining. The relative mRNA expression of type I collagen, type III collagen, and matrix metalloproteinases (MMP-1 and MMP-3) was determined by real-time PCR. In the rat model of photoaged skin, the injected exosomes markedly decreased the epidermal thickness and increased the dermal thickness of the photoaged skin 7 days after treatment. Moreover, the proportion of the stratum corneum of the epidermis was decreased. Furthermore, real-time RT-PCR showed that the mRNA expression of type I collagen was increased and that of type III collagen, MMP-1, and MMP-3 was decreased. Our results demonstrate that ADSC-derived exosome treatment could significantly improve skin photodamage and that ADSC-derived exosomes may be a potential agent for photoaged skin treatment.

Preconditioning With Low-Level Laser Irradiation Enhances the Therapeutic Potential of Human Adipose-derived Stem Cells in a Mouse Model of Photoaged Skin.

This study was conducted to explore the therapeutic potential of human adipose-derived stem cells (ADSCs) irradiated with a low-level laser (LLL). Cultured ADSCs were treated with 650-nm GaAlAs laser irradiation at 2, 4 and 8 J cm-2 . Cell proliferation was quantified by MTT assays, cytokine secretion was determined by enzyme-linked immunosorbent assays, and adipogenic differentiation was examined by oil red O staining. Additionally, the expression profiles of putative ADSC surface markers were analyzed by quantitative real-time PCR. In addition, a mouse photoaged skin model was established by UVB irradiation. Effects of GaAlAs laser-treated ADSCs on the thicknesses of the epidermis and dermis were analyzed by hematoxylin and eosin staining. The results showed that GaAlAs laser treatment of cells at a radiant exposure of 4 J cm-2 enhanced ADSC proliferation and adipogenic differentiation and increased secretion of growth factors. Furthermore, GaAlAs laser irradiation upregulated the expression of putative ADSC surface markers. In the mouse model of photoaged skin, ADSCs treated with GaAlAs laser irradiation had markedly decreased the epidermal thickness and increased the dermal thickness of photoaged mouse skin. Our data indicate that LLL irradiation is an effective biostimulator of ADSCs and might enhance the therapeutic potential of ADSCs for clinical use.

Effect of suction pressures on cell yield and functionality of the adipose-derived stromal vascular fraction.

OBJECTIVE: The present study was performed to explore the effect of suction pressures on the cell yield of adipose-derived stromal vascular fraction (SVF) and the functionality of adipose-derived stem cells (ADSCs), situated in the SVF, and to develop optimal parameters of harvesting SVF for clinical use. METHODS: Adipose tissue was harvested from the lower abdomen of 10 patients by suction-assisted lipoplasty. Suction pressure was either -30 ± 5 kPa or -55 ± 5 kPa. The aspirated samples were subjected to macroscopic observation to verify the adipose particle size and cytological analysis to detect the cell yield and functionality of the SVF harvested. RESULTS: Adipose tissue harvested at -30 ± 5 kPa appeared to have smaller particle sizes and less blood red cells than that harvested at -55 ± 5 kPa. Cell counts revealed that the cell number of the SVF obtained at -30 ± 5 kPa was more than 2-fold higher than that obtained at -55 ± 5 kPa. Cell growth at passages 1 and 2 was faster at -30 ± 5 kPa than that at -55 ± 5 kPa. The secretion of basic fibroblast growth factor and vascular endothelial growth factor as well as the capacity for adipogenic differentiation of the cultured cells at passages 1-3 were higher at -30 ± 5 kPa than those at -55 ± 5 kPa. There was no difference in the expression of the phenotypic markers between the two groups. CONCLUSION: Our data indicate that the pressure for harvesting adipose tissue affects the yield and viability of the SVF. A lower suction pressure is beneficial to harvesting the SVF for clinical use.

Evaluation of 2 Purification Methods for Isolation of Human Adipose-Derived Stem Cells Based on Red Blood Cell Lysis With Ammonium Chloride and Hypotonic Sodium Chloride Solution.

BACKGROUND: The present study was conducted to compare 2 purification methods for isolation of human adipose-derived stromal vascular fraction or stem cells (ADSCs) based on red blood cell (RBC) lysis with 155 mM ammonium chloride (NH4Cl) and hypotonic sodium chloride (NaCl) solution, and try to develop a safe, convenient, and cost-effective purification method for clinical applications. METHODS: Adipose-derived stem cells and RBC were harvested from the fatty and fluid portions of liposuction aspirates, respectively. The suitable concentration of hypotonic NaCl solution on RBC lysis for purification of ADSCs was developed by RBC osmotic fragility test and flow cytometry analysis. The effects of 155 mM NH4Cl or 0.3% NaCl solution on ADSCs proliferation and RBC lysis efficiency were examined by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide assay and lysis efficiency test, respectively. In addition, the adipogenic and osteogenic capabilities, phenotype and genetic stability of ADSCs were evaluated by oil red staining, alkaline phosphatase activity measurement, flow cytometry, and karyotype analysis, respectively. RESULTS: Sodium chloride solution in 0.3% concentration effectively removed RBCs and did not influence the survival of ADSCs in the 10-minute incubation time. The lysis efficiency did not differ significantly between 0.3% NaCl and 155 mM NH4Cl. Moreover, the adipogenic and osteogenic capabilities, surface marker expression and karyotype of the ADSCs were not affected by lysis solutions or by lysis per se. However, the proliferation capacity in the 0.3% NaCl group was superior to that in 155 mM NH4Cl group. CONCLUSIONS: Our data suggest that 0.3% NaCl solution is useful for isolating ADSCs from liposuction aspirate for clinical applications with safety, convenience, and cost-effect.

Primary repair of broad bilateral complete cleft palate with hard palate rotation flap.

It was hard to achieve primary repair for infants with broad bilateral complete cleft palate, and usually multiple procedures are demanded, resulting in multiple injuries and scar hyperplasia, which could influence infants' long-term phonetic function. In this study, primary repair was carried out on 12 cases of 18- to 30-month-old infants who had complete cleft palate by using rotated hard palate flap with a pedicle on the edge of the fissure modified on the basis of traditional von Langenbeck palatoplasty. Eight infants were subjected to initial phonetic sound evaluation at 1 to 2 years after the operation. All the 12 cases of infant complete cheilopalatognathus had fine healing, and no complications such as dyspnea and fistulous opening in palate were observed. The effects of phonetic sound in 8 infants who had this disease were significantly improved as observed in the long-term follow-up. Thus, we consider it as a reliable and feasible procedure to perform hard palate mucosa rotation flap for the primary repair of broad bilateral complete cleft palate. The procedure is simple, while it produces satisfactory therapeutic effects, by which the patients with complete cleft palate could achieve good recovery in their phonetic functions.

[Reconstruction of distal segment defect of thumb with reverse dorsal radial flap].

OBJECTIVE: To introduce the application of reverse dorsal radial flap in reconstruction of distal segment defect of thumb. METHODS: From January 2004 to June 2007, 24 cases of soft tissue defect with exposed bone or tendon at the distal segment of thumb were involved. The flap size ranged from 2 cm x 2 cm to 3 cm x 4 cm. RESULTS: Venous congestion happened in 3 cases with one case of partial necrosis at the distal end of the flap. All the other flaps survived completely. The follow-up period was 6 - 24 months. The appearance and function of the thumbs were satisfactory. The 2-point discrimination was 7 - 11 mm (mean 9.3 mm). CONCLUSIONS: Reverse dorsal radial flap is a good option for reconstruction of distal segment defect of thumb.

[Pudendal-thigh flap: anatomic basis and application in repairing and reconstructing male perineal region].

OBJECTIVE: To investigate the anatomic feature of pudendal-thigh flap and to provide anatomic guide for clinical design and application of the flap in repairing and reconstructing male perineal region. METHODS: The clinical anatomy of blood supply and refluence and nerve innervation of the perineal region in 15 male cadavers (30 sides) were investigated. Then reasonable flaps were designed for male patients needing repair and reconstruction of the perineal regions based on the anatomic findings. RESULTS: Perineal region has adequate arterial blood supply, abundant venous refluence and reliable nerve innervation. The area has multi-source blood supplies: external pudendal artery, anterior cutaneous branches of obturator artery, bole and lateral branch of arteriae scrotales posteriores. The locations of the arteries were mainly uniform in the 30 sides. Three vasoganglion composed by these arteries and their branches connecting superior, central and inferior parts of the flap. In the 23 cases, the largest flap was 17 cm x 9 cm, and the smallest was 8 cm x 5 cm. The flaps survived and recovered well, with perfect appearance and sensation. The patients were all satisfied with the operation results. CONCLUSIONS: The pudendal-thigh flap has abundant blood supply. It can be incised conveniently and covers donor sites well. The flap can be used to repair and reconstruct male perineum with ideal appearance and sensation.

[Expressions of TGF-betaR I, Smad2, Smad3 and Smad7 in keloids].

OBJECTIVE: To investigate the expression of TGF-betaR I, Smad2, Smad3 and Smad7 in keloids, normal scars and normal skins. Discuss the significance of these proteins in the course of keloid. METHODS: Immunohistochemistry method was used to detect the expression intensity and distribution of these proteins in above 3 kinds of different tissues in 44 cases. Statistics was used to analyze the data. RESULTS: The expression of TGF-betaR I were much stronger in keloid than in the other two tissues. The expression of Smad7 were lower in keloids. The increase expression of Smad2,3 were not obvious, but they were found to accumulate in the nucleus. CONCLUSIONS: The results indicate that over-expression of TGF-betaR I, low-expression of Smad7 and accumulation of Smad2,3 may be one of the etiological factors of keloids. This research may provide a new idea to prevent and treat keloids or other fibrosis diseases in the future.

[The effects of chitosan on different-derived fibroblasts].

OBJECTIVE: To investigate the effects of chitosan on the biological activities of the fibroblasts derived from different tissues. METHODS: The biological activities of the fibroblasts derived from different tissues were evaluated with a MTT method for fibroblast proliferation, photic and electronic microscope for morphologic and subcellular structure, 3H-proline uptake method for collagen secretion and ELISA box for the secretion of TGF-beta 1, FGF-AB, and IL-8. RESULTS: This study showed that the chitosan inhabited the proliferation of the fibroblasts and the secretion of the TGF-beta 1, FGF-AB and collagen of the fibroblasts with a dose-depended manner in the normal skin, hypertrophic scar and keloid groups, but it stimulated the IL-8. However, there were no significant differences among the three groups (P > 0.05). CONCLUSION: The chitosan could inhibit the growth, proliferation, biosynthesis and secretion of the fibroblasts, and it may be used to treat different scars.

[Hemodynamic changes of microvascular anastomosis with nitinol clips].

OBJECTIVE: To investigate the hemodynamic changes of the end-to-end anastomosed arteries with nitinol clips. METHODS: Fifteen New Zealand rabbits were divided into anastomosis clip group, suture group and control group randomly. The carotid arteries were resected and end-to-end anastomosis were carried out with nitinol clips in anastomosis clip group and with traditional suture in suture group. The carotid arteries remained undamaged in control group. On the days of 3, 9, 21 and 30 postoperatively, mean blood velocity (Vm), pulsatility index (PI) and resistance index (RI) of anastomosed arteries were determined by Ultrasonography Doppler. RESULTS: On the days of 8 and 9 postoperatively, there were no significant differences of VM, PI and RI between two experimental groups (P > 0.05). On the days of 20 and 30 postoperatively, the differences of Vm and RI were significant (Vm: P < 0.01, P < 0.05: RI: P < 0.01, P < 0.05). The hemodynamic restoration of the anastomosis clip group was better than that of the suture group. CONCLUSION: The hemodynamics of arteries anastomosed with nitinol clips is better than that with traditional suture. This technique has practical value clinically.