Effect of Dermabrasion and ReCell® on Large Superficial Facial Scars Caused by Burn, Trauma and Acnes△.

Objective To explore the effects of dermabrasion combined with ReCell® on large superficial facial scars caused by burn, trauma and acnes.Methods Nineteen patients with large superficial facial scars were treated by the same surgeon with dermabrasion combined with ReCell®. According to the etiology, patients were classified into post-burning group (n=5), post-traumatic group (n=7) and post-acne group (n=7). Fifteen patients completed the follow-ups, 5 patients in each group. Healing time, complication rate, the preoperative and 18-month-post-operative assessments using Patient Satisfaction Score (PSS), Vancouver Scar Scale (VSS), and Patient and Observer Scar Assessment Scale (POSAS) of each group were analyzed to compare the effect of the combined therapy on outcomes.Results The healing time of post-burning group (19.6±4.0 days), post-traumatic group (15.8±2.6 days), and post-acne group (11.4±3.1 days) varied remarkably (F=7.701, P=0.007). The complication rates were 60%, 20%, and 0 respectively. The post-operative POSAS improved significantly in all groups (P<0.05), where the most significant improvement was shown in the post-acne group (P<0.05). The post-operative PSS and VSS improved only in the post-traumatic group and post-acne group (all P<0.05), where the more significant improvement was also shown in the post-acne group (P<0.05).Conclusions The combined treatment of dermabrasion and ReCell® has remarkable effect on acne scars, moderate effect on traumatic scars and is not suggested for burn scars. POSAS should be applied to assess the therapeutic effects of treatments for large irregular scars.

[Autologous Fat Grafting in Scar Revision].

Regenerative medicine is an emerging discipline. Adipose tissue is a rich source of fat cells and mesenchymal stem cells, and autologous fat grafting has increasingly been applied in plastic surgeries and dermatological treatments. This paper reviews the latest advances in autologous fat grafting in scar revision.

[Construction of keratinocyte growth factor phage active peptides for the promotion of epidermal cell proliferation].

OBJECTIVE: To construct and display the keratinocyte growth factor (KGF) phage active peptides so as to detect the promoting effects of epidermal cell. METHODS: KGF sequences were chosen and their primers were designed. The selected genes of P1, P2 and P4 were obtained by reverse transcription (RT)-PCR. P3 was obtained by direct synthesis. And the KGF genes were subcloned into pComb3 vector. The technique of phage display was employed to display the genes on phage surface. Methyl thiazolyl tetrazolium (MTT) assay was used to evaluate the promoting effects of KGF phage active peptides on the proliferation of epidermal cell. Optical density (A) was determined at 570 nm. Immunofluorescent assay was employed to evaluate the cell affinity of KGF phage active peptides. RESULTS: The four KGF genes were obtained and subcloned into pComb3 vector. The proteins of the KGF genes were expressed on the surface of the pComb3 vector. The MTT data of optical density (A) showed that significant differences existed between the negative control and KGF control (0.293 ± 0.017 vs 0.520 ± 0.043) and KGF phage active peptide groups (0.293 ± 0.017 vs 0.469 ± 0.057, 0.441 ± 0.048, 0.438 ± 0.035, 0.446 ± 0.037) (all P < 0.01). The results of immunofluorescent assay indicated that KGF and KGF phage active peptides had excellent cell affinity. CONCLUSION: KGF phage active peptides are successfully constructed and displayed and they may promote the proliferation of epidermal cell.

[The clinical classification method research of keloid].

OBJECTIVE: To explore the clinical classification method of keloids and providing a thread for the treatment of keloids. METHODS: To summarize the 600 cases of keloid patients we accepted and diagnosed from November 2004 to October 2012, and filling in keloid patients information sheet, recording the keloids form by photographs, analyzing the treatment, putting forward the classification method of keloids in clinic. RESULTS: According to the position and quantity that keloids grow, the keloid patients are divided into four major categories:one in single site, one in each site, more than one in single site and more than one in each site; According to the area and thickness of keloids, the keloid single lesion is divided into four subclasses: type of small area and thin, type of small area and thick, type of large areas and thin,type of large areas and thick; According to the number of lesions, keloid multiple lesions is divided into two subgenera: isolated multiple and dispersion multiple, different kinds of keloids suit different methods of treatment. CONCLUSION: The clinical classification method of keloids can be used to provide thought for the treatment of keloids, and have a good application value.

[Skin needle roller importing triamcinolone acetonide into scar to treat hypertrophic scars].

OBJECTIVE: To evaluate the effect of importing triamcinolone acetonide into hypertrophic scars with skin roller needles. METHODS: Thirty-two cases with burn hypertrophic scar were treated. The skin roller needles were moved back and forth on the hypertrophic scars with triamcinolone acetonide dropping on the scar surface at the same time. So the triamcinolone acetonide could be imported into the scar through needles and needle holes. The effect was evaluated as cured, effective, and no effect. The Vancouver scaring criteria and visual analogue scale was used to assess the scar color, thickness, texture and feeling before and after treatment, as well as at the untreated scar area (control). RESULTS: Thirty-two cases were treated 1-3 times, including 28 cases with cured result and 4 cases with effective result. The total effective rate was 100%. The scar color, thickness, texture and feeling was significantly different between the scar before and after treatment, or between the treated and untreated scar (P < 0.05). CONCLUSIONS: Importing triamcinolone acetonide into hypertrophic scars with skin roller needles is effective. It is a new method for the treatment of large hypertrophic scar with medicine.

Transforming growth factor-ß1 phage model peptides isolated from a phage display 7-mer peptide library can inhibit.

BACKGROUND: Transforming growth factor-ß1 (TGF-ß1) is known to have a role in keloid formation through the activation of fibroblasts and the acceleration of collagen deposition. The objective of this current study was to isolate TGF-ß1 phage model peptides from a phage display 7-mer peptide library to evaluate their therapeutic effect on inhibiting the activity of keloid fibroblasts. METHODS: A phage display 7-mer peptide library was screened using monoclonal anti-human TGF-ß1 as the target to obtain specific phages containing ectogenous model peptides similar to TGF-ß1. Enzyme-linked immunosorbent assay (ELISA) was performed to select monoclonal phages with good binding activity, which underwent DNA sequencing. MTT assay and apoptosis assessment were used to evaluate the biological effects of the phage model peptides on keloid fibroblasts. Immunofluorescence assay was employed to show the binding affinity of the model peptides on phages causing keloid fibroblasts. Quantitative real-time PCR analysis was carried out to detect the expressions of nuclear factor κB (NF-κB) mRNA, connective tissue growth factor (CTGF) mRNA and TGF-ß receptor II (TßRII) mRNA in keloid fibroblasts. RESULTS: Specific phages with good results of ELISA were beneficiated. Four phage model peptides were obtained. The data of MTT showed that TGF-ß1 and one phage model peptide (No. 4) could promote keloid fibroblasts proliferation, however, three phage model peptides (No. 1 - 3) could inhibit keloid fibroblasts proliferation. The results of apoptosis assessment showed that the three phage model peptides could slightly induce the apoptosis in keloid fibroblasts. The data of immunofluorescence assay revealed that the model peptides on phages rather than phages could bind to keloid fibroblasts. The findings of quantitative real-time PCR analysis suggested that the expressions of NF-κB mRNA and CTGF mRNA in the three phage model peptide groups decreased, while the expression of TßRII mRNA slightly increased. CONCLUSIONS: Three phage model peptides isolated from a phage display 7-mer peptide library can inhibit keloid fibroblasts proliferation and induce the apoptosis in keloid fibroblasts. They can inhibit the activity of keloid fibroblasts by blocking TGF-ß1 binding to its receptor and then regulating the expressions of NF-κB, CTGF and TßRII.

[Study of TGF-ß1 phage model peptides on inhibiting keloid fibroblasts proliferation].

OBJECTIVE: To isolate the transforming growth factor-beta 1 (TGF-ß1) phage model peptides from phage 12-mer display peptide library to inhibit the proliferation of keloid fibroblasts. METHODS: The phage display 12-mer peptide library was screened for 4 rounds with monoclonal anti-human TGF-ß1 as the target to yield the specific phage model peptides. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used for the quantitative determination of cellular proliferation. Apoptosis was detected by the Annexin V-FITC/PI apoptosis detection kit and the cells were analyzed with flow cytometry. Immunofluorescent assay was employed to show the binding affinity of model peptides for keloid fibroblasts. Quantitative real-time polymerase chain reaction (PCR) was performed to detect the expressions of nuclear factor kappa B (NF-κB) and connective tissue growth factor (CTGF). RESULTS: Ten phage model peptides were obtained and they were similar to TGF-ß1, TGF-ß2, TGF-ß receptor II (TßRII), TGF-ß-induced factor, NF-κB or mitogen-activated protein kinase (MAPK). The results of MTT showed that four phage model peptides (No. 7 - 10) could inhibit the proliferation of keloid fibroblasts (P < 0.05). The results of apoptotic assessment showed that phage model peptides (No. 7 - 10) could slightly trigger the late apoptotic stage of keloid fibroblasts. The data of immunofluorescence assay revealed that the model peptides on phages rather than phages could bind to keloid fibroblasts. The findings of quantitative real-time PCR analysis suggested that the relative expression of NF-κB decreased in phage model peptides groups (No. 7 - 10). The quantitative expression was 0.28, 0.26, 0.46 and 0.30 respectively versus the negative control group. The relative expression of CTGF decreased in phage model peptides groups (No. 7 - 10). The quantitative expression was 0.26, 0.60, 0.34 and 0.17 respectively versus the negative control group. CONCLUSION: Four phage model peptides (No. 7 - 10) isolated from phage display 12-mer peptide library can inhibit the proliferation of keloid fibroblasts via regulating the expressions of NF-κB and CTGF.

[Clinical utility of penile augmentation with prepuce retrocession].

OBJECTIVE: To explore the clinical utility of penile augmentation with prepuce retrocession for redundant prepuce. METHODS: From May 2008 to May 2010, 80 patients of redundant prepuce were surgically corrected by the following steps: injecting physiological saline to make penile erect, sliveing penile inner board, maintaining subcutaneous tissue, fascia, blood vessel, lymphatic ducts and nerve between inner and outer board, pushing and flattening fat and clumsy tissue and keying approximation so as to augment penile diameter. RESULTS: Excellent penile functions were achieved. After a 6-month follow-up, the operative outcome was satisfactory without any complication. The operation with modified circumcision with penile augmentation had the following advantages of a lesser blood loss, mild postoperative edema, no secondary hemorrhage or infection, a quick recovery and a good appearance of penis. CONCLUSION: Treatment of redundant prepuce by penile augmentation with prepuce retrocession is an effective method.

[Effect of survivin antisense oligodeoxynucleotide on proliferation and apoptosis of human malignant melanoma cells].

OBJECTIVE: To investigate the effect of survivin antisense oligodeoxynucleotide (ASODN) on proliferation and apoptosis of human malignant melanoma cells. METHODS: hMMC A375 colonies in log growth phase were collected and divided into control group (C, without transfection), sense chain group [SC, transfected with 600 nmol/L survivin sense oligodeoxynucleotide (ODN)], mismatch chain group (MC, transfected with 600 nmol/L survivin mismatch sense ODN), liposome group (L, treated with liposome), antisense chain group (AC, transfected with survivin ASODN, and subdivided into AC 200, 400, 600 nmol/L subgroups) according to the random number table. Transfection result was observed under inverted fluorescence microscope. Inhibition rate of cell proliferation was calculated after determination of cell viability with MTT method. Cell cycle and apoptosis rate were detected with bi-variable flow cytometry. Expression of survivin protein was determined with Western blot. Activity of caspase-3 was assessed with kinase method. Data were processed with analysis of variance. RESULTS: (1) Cell transfection rates in SC, MC, AC 600 nmol/L groups were all above 80%. (2) Compared with those in SC group [(5.23 +/- 0.25)%], MC group [(5.09 +/- 0.13)%] and L group [(4.70 +/- 0.45)%], inhibition rates of cell proliferation in AC 200, 400, 600 nmol/L groups 24 hours after transfection [(10.30 +/- 0.56)%, (16.69 +/- 0.58)%, (24.67 +/- 0.67)%] were significantly increased (F = 746.91, and P values all below 0.05). As time after transfection went on, proliferation inhibition rate was increased obviously. (3) Apoptosis rate in AC 200, 400, 600 nmol/L groups 24 hours after transfection was respectively (13.5 +/- 1.9)%, (20.1 +/- 1.5)%, (32.1 +/- 2.9)%, which were significantly higher than those in C, SC, MC, and L groups [(6.5 +/- 0.6)%, (5.6 +/- 0.7)%, (6.4 +/- 1.0)%, (6.5 +/- 1.3)%, F = 139.9, P values all below 0.05]. Cells in AC group were blocked in G2/M stage. (4) Compared with those in C group, expression amount of survivin protein decreased, and caspase-3 activity obviously increased (F = 63.1, P values all below 0.05) in AC group. No significant difference in caspase-3 activity between SC, MC, L groups and C group was observed (F = 0.512, P values all above 0.05). CONCLUSIONS: Survivin ASODN can inhibit the proliferation of hMMC A375 in a concentration-time dependent manner, and it induces G2/M stage block and promotes its apoptosis.

Keratinocyte growth factor phage model peptides can promote epidermal cell proliferation without tumorigenic effect.

BACKGROUND: Keratinocyte growth factor (KGF) significantly influences epithelial wound healing. The aim of this study was to isolate KGF phage model peptides from a phage display 7-mer peptide library to evaluate their effect on promoting epidermal cell proliferation. METHODS: A phage display 7-mer peptide library was screened using monoclonal anti-human KGF antibody as the target. Enzyme linked immunosorbent assay (ELISA) was performed to select monoclonal phages with good binding activity. DNA sequencing was done to find the similarities of model peptides. Three-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, immunofluorescence assay and quantitative real-time PCR analysis were employed to evaluate the effect of the phage model peptides on epidermal cells. RESULTS: Thirty-three out of fifty-eight (56.9%) of the isolated monoclonal phages exhibited high binding activity by ELISA. Ten of fifteen obtained phage model peptides were similar to KGF or epidermal growth factor (EGF). MTT assay data showed that four (No. 1 - 4) of the ten phage model peptides could promote epidermal cell proliferation. The expression of keratinocyte growth factor receptor (KGFR) mRNA in the KGF control group and the two phage model peptide groups (No. 1 and No. 2) increased. Expression of c-Fos mRNA and c-Jun mRNA in the KGF control group increased, but did not increase in the four phage model peptide groups (No.1 - 4). CONCLUSION: Four phage model peptides isolated from the phage display 7-mer peptide library can safely promote epidermal cell proliferation without tumorigenic effect.

[Expression and function of sphingosine kinases 1 and 2 in human keloid fibroblasts].

OBJECTIVE: To study the expression and function of sphingosine kinase (SphK) 1 and SphK2 in human keloid fibroblasts. METHODS: Specimens of keloid and surrounding normal skin were collected from 12 patients with keloid during operation. Primary fibroblasts were isolated, cultured, and randomly divided into 3 groups: normal skin group, keloid group, and keloid with transforming growth factor (TGF)-beta1 group cultured with TGF-beta1 for 48 h. Immunofluorescence technique was used to detect the location of SphK1 and SphK2 protein. Real-time PCR and Western blotting were used to measure the mRNA and protein expression levels of SphK1 and SphK2. RESULTS: Sphk1 protein was localized primarily in the nuclei of the fibroblasts, and Sphk2 protein was detected both in the cytoplasm and nuclei in the 3 groups. The mRNA and protein levels of Sphk1 in the keloid group were (0.0608 +/- 0.0190) and (0.8308 +/- 0.1093) respectively, both significantly higher than those of the normal skin group [(0.0383 +/- 0.0147) and (0.6800 +/- 0.1126) respectively, both P < 0.05], but significantly lower than those of the keloid fibroblasts with TGF-beta1 group [(0.0790 +/- 0.0280), P < 0.05, and (1.4267 +/- 0.1938), P < 0.01]. There was no significant differences in the Sphk2 mRNA and protein levels among these 3 groups (all P > 0.05). CONCLUSIONS: Sphk1 plays a leading role in keloid pathogenesis. The SphK1 mRNA and protein levels are increased by TGF-beta1 stimulation in keloid fibroblasts, perhaps indicating that Sphk1 is involved in TGF-beta signal transduction pathway.

[Experimental research of correlation between anatomy structure of rabbit ear and creating hypertrophic scar animal model].

OBJECTIVE: To observe the anatomy structure of rabbit ear and the effect of different operation methods and post-operative treatments on the formation of hypertrophic scar. METHODS: The experimental animals were 25 New Zealand white rabbits. 6 pieces of full skin specimens were obtained from each of the ears in 5 rabbits for histological examination. 6 full-thickness skin wounds (d = 8 mm) were made on different sites of ventral side of each ear in the other 20 rabbits. The total number of the wounds was 240. 120 wounds in 10 rabbits were divided into 4 groups randomly to receive different treatments on day 7 postoperatively. No treatment was performed in the other 120 wounds. The wounds healing and the scar formation were observed for six months. The scars were harvested 4 weeks and 8 weeks after operation for pathologic examination and measurement of scar elevation index (SEI). RESULTS: Histological analysis showed that the anatomy structure was different in different sites of the rabbit ear. The best sites for creating hypertrophic scar model were on the medial margin of the middle- and inferior part of ear. The depth of the wound should reach the cartilage membrane of the ear to facilitate the formation of hypertrophic scar. The second strip crust on day 7 postoperatively enhanced the wounds healing and minimized the scar proliferation and hypertrophy. CONCLUSIONS: There is a close correlation between the anatomy structure of the ear and the creation of hypertrophic scar animal model. The wound site, the depth of wound and the post-operative treatment will affect the formation of hypertrophic scar. The study can help to improve the successful rate of creating hypertrophic scar animal model.

Effect of heparin on production of transforming growth factor (TGF)-beta1 and TGF-beta1 mRNA expression by human normal skin and hyperplastic scar fibroblasts.

Heparin affects both dermal fibroblast proliferation and collagen and may mediate these effects by altering the levels of transforming growth factor-beta1 (TGF-beta1) production and TGF-beta1 mRNA expression as a wound healing modulator. The purpose of this study is to probe the effect of heparin on TGF-beta1 and TGF-beta1 mRNA production by human normal skin and hyperplastic scar fibroblasts. This research investigates the effect of heparin on TGF-beta1 and TGF-beta1 mRNA production by human normal skin and hyperplastic scar fibroblasts with exposure to 0 microg/mL, 100 microg/mL, 300 microg/mL, or 600 microg/mL heparin for 24, 48, 72, or 96 hours in a serum-free in vitro model. Levels of TGF-beta1 in the supernatants and TGF-beta1 mRNA expression of fibroblasts were determined by enzyme-linked immunosorbent assay (ELISA) and real time RT-PCR, respectively. Heparin (300 microg/mL and 600 microg/mL) stimulated TGF-beta1 production by normal skin (26% to 83%) and hyperplastic scar fibroblasts (63% to 85%), with statistical significance (P < 0.05) at various time points. Heparin (300 microg/mL and 600 microg/mL) also stimulated TGF-beta1 mRNA expression by normal skin (12% to 53%) and hyperplastic scar fibroblasts (33% to 52%), with statistical significance (P < 0.05) at various time points. These effects of heparin on normal skin and hyperplastic scar fibroblasts may have implications for hyperplastic scar formation and wound healing in vivo.

[Effects of recombinant adenovirus-mediated double suicide genes on implanted human keloid: experiment with athymic mice].

OBJECTIVE: To detect the effects of the recombinant adenovirus-mediated double suicide genes constructed by Escherichia coli cytosine deaminase (CD)/5-fluorocytosine (5-Fc) and herpes simplex virus-thymidine kinase (HSV-TK)/ganciclovir (GCV)-CDglyTK on implanted human keloids and mechanisms thereof. METHODS: Twenty nude mice were implanted with human keloid obtained during operation so as to establish mouse keloid models and then were randomly divided into 4 equal groups: Group A, injected with normal saline (NS) into the keloid once per 3 days for 18 days totally, Group B injected with NS into the keloid and injected intraperitoneally with 5-Fc and GCV; Group C injected with CDglyTK into the keloid, and Group D injected with CDglyTK into the keloid and 5-Fc and GCV injected intraperitoneally. The volume of the implanted keloid tissue was measured 2, 7, 14, 21, 28, 35, and 42 days after operation. On day 42 the keloid tissues were removed to undergo morphological examination, TUNEL method was used to examine the apoptosis of the fibroblasts, and the expression of Bcl-2 and BAX, products of apoptosis-related genes, were detected by immunohistochemistry. RESULTS: Compared to those before treatment the volume of the implanted keloid of Group D began to decrease since 14 days after treatment time-dependently (all P < 0.05), and the volumes of the other 3 groups continued to increase and peaked on days 21, 14, or 7 respectively (all P < 0.05). Microscopy showed infiltration of a larger quantity of histiocyte in the keloid tissue, and more obvious collagen disorganization and apoptosis of fibroblasts in Group D than in the other 3 groups. The protein expression of Bcl-2 was more remarkable and the protein expression of BAX was less remarkable in Group D than in the other 3 groups. CONCLUSIONS: The recombinant adenovirus-mediated double suicide gene therapy is effective on the implanted keloid tissue. The main mechanism may be induction of apoptosis in the keloid fibroblasts.