Salvianolic Acid B Attenuates Hypertrophic Scar Formation In Vivo and In Vitro.

BACKGROUND: Hypertrophic scars (HTSs) are a fibroproliferative disorder that occur following skin injuries. Salvianolic acid B (Sal-B) is an extractant from Salvia miltiorrhiza that has been reported to ameliorate fibrosis in multiple organs. However, the antifibrotic effect on HTSs remains unclear. This study aimed to determine the antifibrotic effect of Sal-B in vitro and in vivo. METHODS: In vitro, hypertrophic scar-derived fibroblasts (HSFs) were isolated from human HTSs and cultured. HSFs were treated with (0, 10, 50, 100 µmol/L) Sal-B. Cell proliferation and migration were evaluated by EdU, wound healing, and transwell assays. The protein and mRNA levels of TGFßI, Smad2, Smad3, α-SMA, COL1, and COL3 were detected by Western blots and real-time PCR. In vivo, tension stretching devices were fixed on incisions for HTS formation. The induced scars were treated with 100 µL of Sal-B/PBS per day according to the concentration of the group and followed up for 7 or 14 days. The scar condition, collagen deposition, and α-SMA expression were analyzed by gross visual examination, H&E, Masson, picrosirius red staining, and immunofluorescence. RESULTS: In vitro, Sal-B inhibited HSF proliferation, migration, and downregulated the expression of TGFßI, Smad2, Smad3, α-SMA, COL1, and COL3 in HSFs. In vivo, 50 and 100 µmol/L Sal-B significantly reduced scar size in gross and cross-sectional observations, with decreased α-SMA expression and collagen deposition in the tension-induced HTS model. CONCLUSIONS: Our study demonstrated that Sal-B inhibits HSFs proliferation, migration, fibrotic marker expression and attenuates HTS formation in a tension-induced HTS model in vivo. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Salvianolic Acid B Reduces the Inflammation of Fat Grafts by Inhibiting the NF-Kb Signalling Pathway in Macrophages.

BACKGROUND: Autologous fat grafting is a common method for soft tissue defect repair. However, the high absorption rate of transplanted fat is currently a bottleneck in the process. Excessive inflammation is one of the main reasons for poor fat transplantation. Salvianolic acid B (Sal-B) is a herbal medicine that shows promise for improving the effectiveness of fat transplantation. OBJECTIVE: The aim of this study was to improve fat graft survival by injecting Sal-B into fat grafts locally. METHODS: In vivo, 0.2 mL of Coleman fat was transplanted into nude mice along with Sal-B. The grafts were evaluated by histologic analysis at 2, 4, and 12 weeks posttransplantation and by microcomputed tomography at 4 weeks posttransplantation. In vitro ribonucleic acid sequencing, cell proliferation assays, anti-inflammatory activity assays, molecular docking studies, and kinase activity assays were performed in RAW264.7 cells to detect the potential mechanism. RESULTS: Sal-B significantly improved fat graft survival and attenuated adipose tissue fibrosis and inflammation. Sal-B also inhibited the polarization of M1 macrophages in fat grafts. In vitro, Sal-B inhibited the proliferation and activation of inflammatory pathways in RAW264.7 cells. In addition, Sal-B had an inhibitory effect on NF-κB (nuclear factor κ light polypeptide gene enhancer in B cells) signaling. This bioactivity of Sal-B may result from its selective binding to the kinase domain of the inhibitor of NF-κB kinase subunit ß. CONCLUSIONS: Sal-B could serve as a promising agent for improving the effect of fat transplantation by inhibiting the polarization of M1 macrophages through NF-κB signaling.

The effect of glycerol as a cryoprotective agent in the cryopreservation of adipose tissue.

BACKGROUND: Long-term preservation of adipose tissue is crucial for clinical applications. Researchers should consider both efficiency and biosafety when choosing a cryoprotective agent (CPA) for adipose tissue preservation. Glycerol has been applied as a nontoxic CPA for multiple tissues but not adipose tissue. We aimed to evaluate the efficacy of glycerol as a CPA for adipose tissue cryopreservation. METHODS: Fresh human adipose tissues were obtained from patients who underwent liposuction and divided into 1 mL samples. Each sample was randomly mixed with 1 mL of CPA: 60-100% glycerol, 0.25 mol/L trehalose or DMSO + FBS and cryopreserved in - 196 °C liquid nitrogen for one month. After thawing and elution, the tissues were immediately evaluated for activity and structural integrity in vitro. Then, 0.2 mL of each sample was transplanted subdermally to the nude mouse dorsum and harvested after one month for histological examination to assess the effect of the cryopreserved fat in transplantation. RESULTS: After cryopreservation, the samples treated with DMSO + FBS, trehalose, 60% and 70% glycerol had a more integrated structure than the samples in other groups. Tissues preserved with 70% glycerol had the highest G3PDH activity of 24.41 ± 0.70, comparable to 24.76 ± 0.48 in fresh tissue (p > 0.05). Adipose-derived stem cells (ASC) viability, proliferation and differentiation capability were also better preserved in 70% glycerol group. In vivo analysis showed that tissue preserved with 70% glycerol had a retention rate of 52.37 ± 7.53%, significantly higher than other groups. Histological observation demonstrated better structural integrity and viability in 70% glycerol group. Compared to the DMSO + FBS and trehalose groups, the glycerol groups showed lower tissue inflammation. CONCLUSION: Glycerol (70%) is efficient in adipose tissue cryopreservation. Glycerol-based CPAs, which are nontoxic and show biosafety, are a promising solution for clinical tissue cryopreservation.

Salvianolic acid-B improves fat graft survival by promoting proliferation and adipogenesis.

BACKGROUND: Our previous study proved that Salvia miltiorrhiza could enhance fat graft survival by promoting adipogenesis. However, the effect of salvianolic acid B (Sal-B), the most abundant and bioactive water-soluble compound in Salvia miltiorrhiza, on fat graft survival has not yet been investigated. OBJECTIVE: This study aims to investigate whether salvianolic acid B could improve fat graft survival and promote preadipocyte differentiation. The underlying mechanism has also been studied. METHODS: In vivo, 0.2 ml of Coleman fat was transplanted into nude mice with salvianolic acid B. The grafts were evaluated by HE and IF at 2 and 4 weeks posttransplantation and by micro-CT at 4 weeks posttransplantation. In vitro, the adipogenesis and proliferative activities of salvianolic acid B were analyzed in cultured human adipose-derived stem cells (h-ADSCs) and 3T3-L1 cells to detect the mechanism by which salvianolic acid B affects graft survival. RESULTS: In vivo, the weights and volumes of the fat grafts in the Sal-B-treated groups were significantly higher than those of the fat grafts in the control group. In addition, higher fat integrity and more viable adipocytes were observed in the Sal-B-treated groups. In vitro, salvianolic acid B showed the ability to promote 3T3-L1 and h-ADSC proliferation and adipogenesis. CONCLUSIONS: Our in vitro experiments demonstrated that salvianolic acid B can promote the proliferation of adipose stem cells and enhance the differentiation of adipose stem cells. Simultaneously, in vivo experiments showed that salvianolic acid B can improve the survival rate of fat transplantation. Therefore, our research shed light on the potential therapeutic usage of salvianolic acid B in improving the survival rate of fat transplantation.

[The significance of ultrasound and mammography in the diagnosis of fat necrosis after fat transplantation in breasts].

OBJECTIVE: To investigate the significance and images characters of ultrasound and mammography in the diagnosis of fat necrosis after fat transplantation in breasts(FTB). METHODS: From June 2005 to June 2012, 13 cases with 96 fat necrosis lumps after FTB, were retrospectively reviewed. The pre-operative images of ultrasound and mammography, as well as the post-operative histologic results were collected. RESULTS: All the 96 lumps were histologically diagnosed as fat necrosis. They were lipo-necrotic cysts with thin wall(29 lumps), or thick wall(52 lumps), or no wall(15 lumps). The cysts with wall could be distinguished both in the images of ultrasound and mammography, but the cysts without wall could only be found in mammography images, not in ultrasound images. CONCLUSIONS: Most fat necrosis lesions show a special cystic images with thin or thick wall in ultrasound and mammography. Irregular lesions without wall can be distinguished in the mammography images, but may be mis-diagnosed in the images of ultrasound. The images characters of fat necrosis lesions in mammography are not confused with the breast tumor.

Treatment of breast injection with polyacrylamide hydrogel with infiltrated fascia capsule removal: report on 104 cases.

BACKGROUND: Complications of polyacrylamide hydrogel injection for augmentation mammaplasty have captured the attention of both medical experts and the general public in China. Increasing numbers of patients are seeking to have the polyacrylamide gel removed from their breasts regardless of whether they suffer from symptoms. Blunt aspiration, which is widely performed but removes only free hydrogel, leaves residual hydrogel in the breast. We report and discuss our ideas and techniques for the treatment of polyacrylamide hydrogel-injected breasts. METHODS: Between February 2005 and December 2010, 104 women who had undergone injection of polyacrylamide hydrogel for breast augmentation presented to us for treatment. A periareolar incision was made on the lower border of the areola, followed by dissection to the outer surface of the infiltrated fascia of polyacrylamide hydrogel. The fascia was dissected away as completely as possible and the free polyacrylamide hydrogel was removed from the cavity. RESULTS: All of the patients were satisfied with the surgical results, except for three patients who complained about the flat appearance of the breast. Preoperative symptoms such as breast lumps, pain, uncomfortable upper-extremity movement, and breast firmness were relieved and disappeared after removal of the free polyacrylamide hydrogel and infiltrated capsule. CONCLUSION: The removal of as much of the infiltrated capsule and free polyacrylamide hydrogel as possible may be successful in removing breast lumps, pain, and uncomfortable upper-extremity movement due to breast firmness. To remove as much polyacrylamide hydrogel as possible, the removal of the infiltrated fascia and capsule together with the free polyacrylamide hydrogel is recommended. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article.

[Integrated autologous fat graft in face recontouring].

OBJECTIVE: To discuss the integrated autologous fat graft technique in face recontouring. METHODS: In this study we treated 83 cases of face recontouring with 3L3M technique (low pressure suction, low speed centrifugation, low volume, multi-plane, multi-tunnel, multi-point injection). Each case was treated 1-3 times and the interval period is 3-6 months. The result was based on comparing the photos taken from pre-operation and post operation, observing the expression recovery, cysts, local absorption, and patients self evaluation. RESULTS: Long time follow up showed that fat graft can be alive in the recipient site for long time after 1-3 times autologous fat injection. More than 73.5% patients were satisfactory with the curative effect while less than 4.8% patients were unsatisfactory. CONCLUSION: 3L3M integrated fat graft technique is an effective and safe treatment in face recontouring.

An integrated fat grafting technique for cosmetic facial contouring.

BACKGROUND: Although several methods of fat grafting have been reported, there is no agreement regarding which technique should be used for autologous fat grafting and the final outcome after fat grafting often depends on the technique used by a surgeon. In this report we present an integrated fat grafting technique developed over the years for cosmetic facial contouring. METHOD: Fat grafts were harvested under low pressure with syringes, processed with low speed centrifugation, and then injected into the affected areas of the face with low volume for each pass via multi-tunnels, multi-planes, and multi-points. We describe the above integrated fat grafting technique as '3Ls and 3Ms' technique. A total of 83 patients with temporal, cheek, and periocular hollowing, lean or aging face or facial asymmetry were injected with autologous fats between 1 and 3 times at interval of 3 to 6 months. All patients were evaluated clinically and followed up to 8 years. RESULTS: The absorption was found variable in different patients. Long-term follow-up proved that final correction after 1-3 injections of autologous fat had persisted for many years. Obviously improved facial contour was evident in most patients after autologous fat transplantations. More than 73.5% of the patients in this series were assessed as satisfactory by all 3 observers (patient, surgeon, and layerperson). Between 12.0% and 21.7% of the patients were mostly satisfactory. Less than 4.8% of the patients were unsatisfactory. CONCLUSIONS: Autologous fat grafting to the face is a safe and reliable option to improve facial contour in patients. The transplanted fat can survive well in the face with our integrated fat grafting technique. More than one injection may be required to achieve optimal outcome.

Autologous fat grafting to the breast for cosmetic enhancement: experience in 66 patients with long-term follow up.

BACKGROUND: Autologous fat grafting to the breast for cosmetic enhancement remains controversial because the efficacy and fate of fat grafting to the breast are primarily unknown. In this report, we present our retrospective study in 66 patients who underwent autologous fat grafting to the breast for various cosmetic reasons and who were followed with sonography, mammography, or magnetic resonance imaging (MRI). METHODS: Sixty-six patients who desired cosmetic enhancement of the breast for various reasons underwent autologous fat transplantation between August 2000 and March 2005 in our institution. The cosmetic outcome was assessed by the plastic surgeons as well as the patients. The imaging features of fat necrosis, cyst formation, and calcification in these patients were carefully studied and biopsies of palpable lumps were evaluated histologically. RESULTS: All patients were followed from 13 to 61 months with an average of 37 months. Breast cosmetic contour was significantly improved in 28 patients (42.4%), improved in 24 patients (36.4%), and not improved in 14 patients (21.2%) as judged by the plastic surgeons. Twenty-seven patients (40.9%) were very satisfied, 26 patients (39.4%) were satisfied, and 13 patients (19.7%) were unsatisfied. Eleven patients (16.7%) developed liponecrotic cysts but only two patients elected to have the breast lump surgically removed. CONCLUSION: Autologous fat grafting to the breast can be a useful procedure for cosmetic enhancement in many patients who desire such a procedure. Patients with breast contour deformities after removal of silicon implants were found to be the best candidates for fat grafting. The primary long-term complication is the formation of liponecrotic cysts which have characteristically benign appearances in sonography, mammography or MRI.

[Three case reports of abdominal compartment syndrome after full abdominoplasty].

OBJECTIVE: To improve the safety of the abdominoplasty by the study of the complication of abdominal compartment syndrome after abdominoplasty. METHODS: Three cases were analyzed and discussed respectively in aspects of clinic symptoms, pathological reason, treatments and follow-up results. The treatment was the incision of musculoaponeurotic system, which usually cause defect of abdominal wall after pressure releasing. The defect of musculoaponeurotic system was reconstructed by Mesh, and the defect of skin was repaired by skin graft. RESULTS: The pathologic change of the complications was the shrinking of the belly cavity's volume and the increasing internal pressure of the abdomen caused the internal pressure of the thoracic cavity increase. The clinical manifestations included compressive feelings of chest and abdomen, high urinary frequency and psychiatric symptom. 2 patients accepted the operation. The abdominal compartment syndrome disappeared after the operation, and the results of the two years follow-up were satisfied. CONCLUSIONS: Full abdominoplasty has the risk of causing abdominal compartment syndrome. This complication can be cured by decompression of abdominal wall. The effect is stable after long time's follow-up. Special preventive methods should be used in high-risk patients to avoid this complication.

[Application of aesthetic sub-unit principle in nasal reconstruction and repairing of nasal defects].

OBJECTIVE: To evaluate the significance and effect of nasal aesthetic sub-unit principle. METHODS: Based on nasal aesthetic sub-unit, 27 patients were divided into 5 groups according to their nasal defects. They were nasal dorsum, tip, alar lobule, soft triangle and two or two above nasal subunit defects groups. Local restore were applied in defects which were less than 50 percent of one nasal subunit. Total nasal subunit reconstruction were applied, if areas of defects were larger than 50 percent of one nasal subunit 10 cases were received free tissue transplantation (including free skin transplantation, composite auricle transplantation, etc), 4 cases were received local flap, and 13 cases were received forehead flap. Following up for 5 to 37 months, evaluation item were contour and color. RESULTS: Skin transplantation and local flap were all survived. Auricle composite flap were all survived except for 2 cases whose epidermis looked red and purple. These two case's epidermis regained normal 6 months after operation. Forehead flap for nasal reconstruction were all survived, but one of them had to perform secondary surgery for coverage of the exposed nasal frame. 27 cases were followed up in all. Patients and their family members were satisfied with the nasal's contour and color. CONCLUSIONS: Good aesthetic effects comes from nasal aesthetic sub-unit principle in nasal reconstruction and repairing of nasal defects.

[Research of the preservation of the composite facial allograft].

OBJECTIVE: To investigate the effective method of preserving composite facial allograft so as to attenuate ischemic injury. METHODS: The composite facial allografts were harvested from dog, perfused and preserved with 4 degrees C physiological sodium chloride and UW solution respectively. Immediately after the removal of the flap, after 12, 24, 48 h of preservation, MTT assay was used to determine the viability of several kinds of tissue, including skin, mucosa, muscle, bleed vessel, nerve and gland. The results of the two groups were compared in term of viability percentage. The pathology of several tissues were observed after 24 and 48 h of storage. RESULTS: The viability percentage of every tissue conserved in UW solution for 48 hours was more than 75%. There was significant difference between physiological sodium chloride group and UW group (P < 0.05). Some changes, including Porous arrangement of fibers in connective tissue of skin and mucosa, hyalinization of tissue around the hair follicle and edema of cell in hair follicle, enlargement of space between muscle bundles and unclearness of boundary of acinus could be seen in physiological sodium chloride group while no significant change in UW group. CONCLUSIONS: UW solution could be considered as preservation solution for composite facial allograft.

[Establishment of composite facial and scalp allograft transplantation model in canine].

OBJECTIVE: To develop an experimental model of composite facial and scalp allograft in canine in order to investigate technical and immunological aspects and functional recovery of facial muscles of this new approach to facial reconstruction. METHODS: (1) Anatomic study: Four mongrel dogs were used for anatomical dissection of the head and neck region and for harvesting flap experiment. (2) Autologous transplantation (group I): Three types composite facial and scalp autologous transplantation were performed in five mongrel dogs. Type I composite tissue flap (group I a n = 2) included bilateral external ear and orbicularis oculi muscle. Type II (group I b n = 1) included single-lateral external ear, orbicularis oculi muscle, external nose upper and lower lip. Type III (group I c n = 2) included single - lateral external ear and orbicularis oculi muscle. (3) Allograft transplantation (group II): In group II a (n = 2), two allograft transplantation were performed with type III composite facial and scalp . In group II b (n = 4), four allograft transplantation were performed with the modified type III composite facial and scalp which included single - lateral external ear, orbicularis oculi muscle and one third of inferior tarsal plate and palpebral conjunctiva. To prevent allograft rejection, Cyclosporin A (CsA) and Methylprednisolone (MP) or Prednisone (PS ) were combined used as immunosuppressive protocol . Dose of CsA was adjusted depending on its blood drug level. Electromyogram (EMG) of orbicularis oculi muscle was carried out at 4 weeks, 6 weeks, 12 weeks and 6 months postoperation. RESULTS: (1) The facial anatomic characteristic of dog is similar to that of human being, external carotid artery and external jugular vein afford good blood supply to composite facial and scalp. (2) The dogs in group I c were long-term surviving with leakage of salivary juice. (3) In group II a (n = 2), one dog presented rejection reaction at 28th day postoperation, the reversal of rejection was achieved by increasing the dose of CsA and prednisone and with topical clobetasol for 2 weeks, the dog survived indefinitely( > 309 days). In group II b (n = 4), there were three dogs survived indefinitely ( > 159 days, > 129 days, > 108 days) without complication, EMG showed the function of orbicularis oculi muscle was gradually improving. CONCLUSION: The modified type III composite facial and scalp allograft transplantation model is an ideal model for facial allograft transplantation study.

[Nasal reconstruction with forehead skin flap and muscle flap].

OBJECTIVE: To improve the paramedian forehead flap for nasal reconstruction. METHODS: Based on the findings of the Cutaneous branch of supratrochlear artery, the forehead musculo-cutaneous flap was divided into the musculo-flap and the skin-flap in 3 patients. The musculo-flap were used to reconstruct the septi-bone structure, and envelop silicon nasal-frame, while the skin-flap were used to reconstruct the nose. In 6 patients, the forehead flap was used as a skin-flap just with the muscle pedicle to reconstruct a nose with good appearance of nasal subunits, without the secondary operation. RESULTS: In all of 9 cases, the cutaneous-branch of supratrochlear artery was found 1.7-2.0 cm above the orbital-rim. Eight patients got good results, only 1 patient had to do the secondary operation to coverage the exposed nasal frame. CONCLUSIONS: The cutaneous-branch of supratrochlear artery could be an anatomical finding to form the forehead skin-flap and it is a good choice to be used to reconstruct a nose, even in complicated cases.