Unsatisfactory Upper Blepharoplasty Revision Technique: Reconstruction of the Upper Orbital Septum and its Fusion Point.

BACKGROUND: The orbital septum prevents both superficial eyelid infection and the spread of bleeding into the orbit. The fusion point between the upper eyelid orbital septum and the levator aponeurosis or the tarsal plate plays an important role in determining the width of the natural double eyelid. We observed a number of unsatisfactory upper eyelid blepharoplasty outcomes resulting from improper handling of the orbital septum-such as excessive destruction of orbital septum tissue and failure to form a firm attachment point between the orbital septum and the levator aponeurosis or palpebrae plate-during primary surgery. OBJECTIVES: The three most common types of unsatisfactory upper blepharoplasty outcomes include abnormally high double eyelid creases, multiple creases, and disappearance of creases. In the repair operation, we try to determine the remaining orbital septum tissue for reconstruction and form a firm attachment between the orbital septum and the levator aponeurosis or tarsal plate. Follow-up after surgery was performed to observe whether our technique can ensure effective and favorable long-term natural-looking upper eyelid blepharoplasty outcome. METHODS: From January 2018 to January 2020, secondary blepharoplasty involving the above-mentioned unsatisfactory double eyelid results was performed in 83 patients, including 63 patients (141 eyes) with abnormally high skin creases, 6 patients (8 eyes) with multiple creases, and 14 patients (24 eyes) with double eyelid disappearance. The outcomes were assessed 6 months to 2 years after the surgery by reviewing the photographs to evaluate the esthetic outcomes including stability of double eyelid, double fold curve, symmetry, patient satisfaction, and the incidence of complications. RESULTS: After an average follow-up of 12 months, most patients achieved a better double eyelid appearance. The esthetic outcome was graded as good in 80 patients, poor due to recurrence of double eyelid disappearance in 2, and poor because of asymmetry of the double eyelid curve in length or width in 1 patient. All patients had acceptable scars. No cases of infection or ptosis were observed. CONCLUSION: Reconstructing the orbital septum and ensuring a firm fixation with the levator aponeurosis or tarsal plate is an effective method to repair unsatisfactory upper eyelid blepharoplasty. Moreover, it is very important to protect the orbital septum and proper treatment during the initial surgery. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. 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 .

Construction of a dermis-fat composite in vivo: Optimizing heterogeneous acellular dermal matrix with in vitro pretreatment.

Dermis-fat composite tissues have been widely used in plastic and reconstructive surgery and were previously constructed using hydrogel-type scaffolds. The constructs can be used for in vitro cosmetic and pharmaceutical testing but are not mechanically strong enough for in vivo applications. In this study, we used heterogeneous (porcine) acellular dermal matrix (PADM) as dermal layer scaffold. PADM was pretreated with the laser micropore technique and then precultured with rat adipose-derived stem cells (rADSCs) in vitro. rADSCs proliferated well on pretreated/unpretreated PADM, showing increased expression of genes associated with inflammatory regulation, proangiogenesis, and stemness, indicating that pretreated/unpretreated PADM both provide a beneficial microenvironment for rADSCs to exert their paracrine function. After in vitro processing, the rADSCs-polyporous PADM and PADM without pretreatments were implanted into the back of rats respectively, followed by adipose tissue transplantation. After implantation, the inflammation induced by pretreated PADM was significantly attenuated and localized compared to the unpretreated group. Moreover, the vascularization was faster, and more adipose tissue was formed in the pretreated group. Sound dermis-fat composite tissue was constructed with sufficient strength, which can potentially be used for actual repair application.

Pre-culture of adipose-derived stem cells and heterologous acellular dermal matrix: paracrine functions promote post-implantation neovascularization and attenuate inflammatory response.

Heterologous acellular dermal matrix (ADM) has good biocompatibility and sufficient strength for clinical use for the repair of defects, tissue filling, and resurfacing of deep wounds. However, ADM tissue has such a compact structure that it can easily result in delayed vascularization after implantation. Moreover, in spite of the low immunogenicity of heterologous ADM, it can still cause varying degrees of inflammation in the host. These two drawbacks limit the efficacy and scope of clinical applications for heterologous ADM. Adipose-derived stem cells (ADSCs) have multiple effects on promoting vascularization and regulating immunological responses through paracrine signaling. Pre-culturing heterologous ADM with ADSCs may address these problems; however, it is unknown if ADSCs can exert their paracrine functions within a heterologous ADM microenvironment. This study examined the effect of porcine ADM (PADM) on the paracrine function of rat ADSCs (rADSCs) and showed that the expression of genes associated with inflammatory regulation, pro-angiogenesis factors, and stemness increased when rADSCs were seeded on PADM compared to rADSCs seeded on microplates. This indicates that PADM can provide a beneficial microenvironment for ADSCs to exert their paracrine function. After pre-culture, in vivo implanted rADSC-PADM exhibited improved vascularization and mitigated inflammatory response compared to untreated PADM. This study is the first to report that ADM can provide a suitable microenvironment for ADSCs and that pre-culturing improved the ADM implantation quality in vivo. These results suggest that it could be possible to apply heterologous ADM more effectively and broadly for repair and reconstruction treatments.

In vitro proliferation and differentiation of adipose-derived stem cells isolated using anti-CD105 magnetic beads.

The present study aimed to investigate the feasibility of isolating adipose-derived stem cells (ADSCs) by selecting cells that express the surface receptor CD105. Surface antigen expression of the unsorted cells was undertaken using FACS analysis. Primary adipose-derived cells were isolated. The second passage cells were incubated with anti-CD105 magnetic beads, and separated using a magnetic separator. Cell growth and colony formation was determined by counting and Giemsa staining, respectively. Cells also underwent histological immunohistochemical, and RT-PCR analyses to determine their chondrogenic, adipogenic and osteogenic potential. Increased cell proliferation and colony formation was observed in CD105-positive (CD105⁺) as compared to the CD105-negative (CD105⁻) cells (P<0.001). Following induction, the expression of type II collagen and the number of calcium deposits and lipid droplets in the CD105⁺ ADCs were markedly higher than in the CD105⁻ ADCs. Furthermore, increased alkaline phosphatase (AKP), leptin and PPARγ2 mRNA expression was detected in the CD105⁺ ADCs (P<0.01). Isolation of CD105⁺ ADSCs by MACS was feasible. Thus, CD105 can be used as a relatively specific marker for the selection of ADSCs. Although the chondrogenic, adipogenic and osteogenic potential of these cells is suggestive of their potential for use in tissue engineering treatments, further in vivo studies are necessary.

A modified technique combining excision of the levator muscle and tarsus for blepharoptosis in Asians: a 6-year experience with 116 cases.

BACKGROUND: In patients with blepharoptosis, the function of levator muscle is insufficient or completely absent, causing blepharoptosis in various degrees. For mild or moderate blepharoptosis, levator advancement or resection is commonly performed. However, in severe cases, undercorrection results and recurrence often occur even a great length of levator muscle is resected. Because the levator muscle makes the upper eyelid move in a physiologic direction, exerting the function of residual levator muscle is still a more preferred approach for correction of blepharoptosis. This study combined tarsus resection with levator resection. The resected tarsus can offset the amount of the levator excised, making this technique applicable for severe cases. METHODS: This study included 116 patients (175 eyelids) with moderate or severe ptosis who underwent combined excision of the levator muscle and the tarsus. For cases of bilateral blepharoptosis with different levator functions between the two eyelids, surgery was performed for more severe side first and for the other side 6 months later. Postoperatively, the correction and symmetry results were evaluated and analyzed using chi-square testing by SPSS (version 10.0). RESULTS: Adequate or normal correction was achieved in 149 eyelids (85.1%). The difference in correction results did not differ significantly between moderate and severe cases. With a two-stage operation, 98 patients (84.5%) obtained good or fair asymmetry results, and no statistically significant difference existed between the bilateral and unilateral cases. CONCLUSION: The described technique appears to be effective for both moderate and severe ptosis, with better biomechanics and a satisfying aesthetic outcome.

Using human hair follicle-derived keratinocytes and melanocytes for constructing pigmented tissue-engineered skin.

BACKGROUND: Traditional tissue-engineered skin does not produce a satisfactory long-term result because it lacks natural skin pigmentation and leads to discolored cosmetically unpleasing skin that only functions to cover the body of patients. Additionally, the cell sources for tissue-engineered skin are generally derived from normal skin, which is often limited in patients with skin defects. METHODS: In this study, hair follicle melanocytes and keratinocytes were isolated from human scalp. The melanocytes were co-cultured with keratinocytes until the second passage and then purified. Purified melanocytes and keratinocytes were seeded onto the chitosan-gelatin membrane for 1 week to construct pigmented tissue-engineered skin. The pigmented skin equivalent was used to resurface the skin defect in nude mice. Four weeks after grafting, skin biopsies were harvested to take hematoxylin and eosin staining and immunohistochemistry staining of Melan-A and HLA-ABC. RESULTS: Large quantities of purified melanocytes can be obtained with co-culture method. The hematoxylin and eosin staining of repaired skin biopsy demonstrated that the tissue-engineered skin can repair skin defects successfully. Engineered skin contained pigmentation and stained positive for Melan-A and HLA-ABC, which confirmed the presence of melanocytes and its sources were of human origin. CONCLUSION: This study demonstrated the possibility of constructing pigmented tissue-engineered skin with human hair follicle-derived keratinocytes and melanocytes, which brings a promising method to make up for the deficiency of traditional tissue-engineered skin and provides an alternative treatment for depigmentation diseases.

[Preliminary study of constructing tissue-engineered cartilage with the endoskeletal scaffold of HDPE by bone marrow stromal cells].

OBJECTIVE: To explore the feasibility of using a nonreactive, permanent endoskeletal scaffold to create the prothesis in special shape which is covered with tissue-engineered cartilage. METHODS: Porcine BMSCs and articular chondrocytes were isolated and expanded respectively in vitro. Porcine BMSC of passage 1 in the concentration of 10 x 10(7)/ml were seeded onto a cylinder-shaped PGA (1 mm in thickness)/Medpor (3mm in diameter and 5mm in highness) scaffold as the experimental group. After the cell-scaffold constructs were cultured for 5 days, the primary medium, high-glucose DMEM medium with 10% fetal bovine serum (FBS), was replaced by chondrogenically inductive medium for 4 weeks. BMSCs and chondrocytes of the same concentration were seeded respectively onto the scaffold as the negative control group and the positive control group. After cultured in vitro for 4 weeks, the cell-scaffolds construct were implanted into subcutaneous pockets on the back of nude mice. Four and eight weeks later, the formed cartilage prosthesis were harvested and then evaluated by gross view, histology, immunohistochemistry and glycosamino-glycan (GAG) content. RESULTS: Cells in all groups had fine adhesion to the scaffold and could secrete extracellular matrix. All specimens in experimental group and positive control group formed mature cartilage with collagen II expression.The mature catrtilage wraped HDPE compactly and grown into the gap of HDPE. Mature lacuna structures and metachromatic matrices were also observed in these specimens. GAG contents in experimental group were (5.13 +/- 0.32) mg/g (4 weeks), (5.37 +/- 0.12) mg/g (8 weeks). In contrast, specimens in BMSC group showed mainly fibrous tissue. CONCLUSION: It indicates that it is feasible to create special shaped tissue-engineering cartilage with the permanent internal support using BMSCs as seed cell.

[Creation of tissue engineered cartilage with internal support].

OBJECTIVE: To test the hypothesis that tissue-engineered cartilage can be bioincorporated with a nonreactive, permanent endoskeletal scaffold. METHODS: Chondrocytes obtained from swine articular were seeded onto polyglycolic acids(PGA) scaffold which was incorporated with high-density polyethylene (Medpor). After cultured in vitro for two weeks,the cell-scaffold construct was implanted into subcutaneous pockets on the back of nude mice. Six weeks later,the newly formed cartilage prosthesis was harvested, and a small part of sample was evaluated by gross view, histology, type II collagen immunohistochemistry and biochemistry. PGA scaffold seeded with cells as the control group. RESULTS: The newly formed cartilage was very similar to normal cartilage in both gross view and histology, and jointed Medpor tightly. The center of control group was hollow. CONCLUSION: This pilot technique combining tissue engineering with a permanent success in creating cartilage without "hollow" phenomenon. biocompatible endoskeleton demonstrated

[Effects of nandrolone phenylpropionate on the expression level of pro alpha 1 (I) collagen after burn in rats].

OBJECTIVE: To explore the effects of nandrolone phenylpropionate (NP) on the expression level of pro alpha 1 (I) collagen after burn in rats and the possible mechanism involved in the process. METHODS: Thirty-two Wistar rats with a deep second-degree scald injury and 20% of total body surface area were randomly divided into two groups to receive either 5 mg/kg NP(NP group) or normal saline (control group) every other day. We analyzed the mean integrated optical density(mIOD) of androgen receptor (AR) to determine the distribution and expression of AR in fibroblasts by immunohistochemistry, and measured expression level of pro alpha 1 (I) collagen mRNA by quantitative fluorescent RT-PCR to find the relation between expressions of AR and pro alpha 1 (I) collagen mRNA. The total specimens were obtained from the scalded rats after 4, 7, 14 and 21 of after burn. RESULTS: The expression of pro alpha 1 (I) collagen mRNA in NP group was significantly higher than that in control group on the 7th, 14th and 21st days(P < 0.05), but there was no significant difference on the 4th day. The density of AR in fibroblasts had significant difference (P < 0.05) between the two groups after 4, 7, 14 and 21 days. A positive relationship existed between the expression of pro alpha 1 (I) collagen mRNA and quantity of AR in fibroblasts(r = 0.836). CONCLUSION: The nandrolone phenylpropionate increased the expression of pro alpha 1 (I) collagen mRNA and enhanced the density of AR in fibroblasts. The higher expression of pro alpha 1 (I) collagen mRNA had a relation with the change of quantity of AR in fibroblasts.