The regenerative role of adipose-derived stem cells (ADSC) in plastic and reconstructive surgery.

The potential use of stem cell-based therapies for the repair and regeneration of various tissues and organs offers a paradigm shift in plastic and reconstructive surgery. The use of either embryonic stem cells (ESC) or induced pluripotent stem cells (iPSC) in clinical situations is limited because of regulations and ethical considerations even though these cells are theoretically highly beneficial. Adult mesenchymal stem cells appear to be an ideal stem cell population for practical regenerative medicine. Among these cells, adipose-derived stem cells (ADSC) have the potential to differentiate the mesenchymal, ectodermal and endodermal lineages and are easy to harvest. Additionally, adipose tissue yields a high number of ADSC per volume of tissue. Based on this background knowledge, the purpose of this review is to summarise and describe the proliferation and differentiation capacities of ADSC together with current preclinical data regarding the use of ADSC as regenerative tools in plastic and reconstructive surgery.

Nerve regeneration with aid of nanotechnology and cellular engineering.

Repairing nerve defects with large gaps remains one of the most operative challenges for surgeons. Incomplete recovery from peripheral nerve injuries can produce a diversity of negative outcomes, including numbness, impairment of sensory or motor function, possibility of developing chronic pain, and devastating permanent disability. In the last few years, numerous microsurgical techniques, such as coaptation, nerve autograft, and different biological or polymeric nerve conduits, have been developed to reconstruct a long segment of damaged peripheral nerve. A few of these techniques are promising and have become popular among surgeons. Advancements in the field of tissue engineering have led to development of synthetic nerve conduits as an alternative for the nerve autograft technique, which is the current practice to bridge nerve defects with gaps larger than 30 mm. However, to date, despite significant progress in this field, no material has been found to be an ideal alternative to the nerve autograft. This article briefly reviews major up-to-date published studies using different materials as an alternative to the nerve autograft to bridge peripheral nerve gaps in an attempt to assess their ability to support and enhance nerve regeneration and their prospective drawbacks, and also highlights the promising hope for nerve regeneration with the next generation of nerve conduits, which has been significantly enhanced with the tissue engineering approach, especially with the aid of nanotechnology in development of the three-dimensional scaffold. The goal is to determine potential alternatives for nerve regeneration and repair that are simply and directly applicable in clinical conditions.

Screening of common CYP1B1 mutations in Iranian POAG patients using a microarray-based PrASE protocol.

PURPOSE: The gene coding cytochrome P4501B1 (CYP1B1) has been shown to be a major cause of primary congenital glaucoma in the Iranian population. More recently it was shown to also be important in juvenile-onset open angle glaucoma (JOAG). We aimed to further investigate the role of CYP1B1 in a larger cohort of primary open angle glaucoma (POAG) patients which included late-onset patients. We also aimed to set up a microarray based protocol for mutation screening with an intent of using the protocol in a future population level screening program. METHODS: Sixty three POAG patients, nine affected family members, and thirty three previously genotyped primary congenital glaucoma (PCG) patients were included in the study. Clinical examination included slit lamp biomicroscopy, IOP measurement, gonioscopic evaluation, fundus examination, and measurement of perimetry. G61E, R368H, R390H, and R469W were screened by a protocol that included multiplexed allele specific amplification in the presence of a protease (PrASE), use of sequence tagged primers, and hybridization to generic arrays on microarray slides. The entire coding sequences of CYP1B1 and myocilin (MYOC) genes were sequenced in all individuals assessed by the microarray assay to carry a mutation. Intragenic single nucleotide polymorphism (SNP) haplotpes were determined for mutated alleles. RESULTS: Genotypes assessed by the array-based PrASE methodology were in 100% concordance with sequencing results. Seven mutation carrying POAG patients (11.1%) were identified, and their distribution was quite skewed between the juvenile-onset individuals (5/21) as compared to late-onset cases (2/42). Four of the seven mutation carrying Iranian patients harbored two mutated alleles. CYP1B1 mutated alleles in Iranian PCG and POAG patients shared common haplotypes. MYOC mutations were not observed in any of the patients. CONCLUSIONS: The PrASE approach allowed reliable simultaneous genotyping of many individuals. It can be an appropriate tool for screening common mutations in large sample sizes. The results suggest that CYP1B1 is implicated in POAG among Iranians, notably in the juvenile-onset form. Contrary to POAG patients studied in other populations, many mutation harboring Iranian patients carry two mutated alleles. We propose an explanation for this observation.

Nanotechnology and bio-functionalisation for peripheral nerve regeneration.

There is a high clinical demand for new smart biomaterials, which stimulate neuronal cell proliferation, migration and increase cell-material interaction to facilitate nerve regeneration across these critical-sized defects. This article briefly reviews several up-to-date published studies using Arginine-Glycine-Aspartic acid peptide sequence, nanocomposite based on polyhedral oligomeric silsesquioxane nanoparticle and nanofibrous scaffolds as promising strategies to enhance peripheral nerve regeneration by influencing cellular behaviour such as attachment, spreading and proliferation. The aim is to establish the potent manipulations, which are simple and easy to employ in the clinical conditions for nerve regeneration and repair.

Investigation of Schwann cell behaviour on RGD-functionalised bioabsorbable nanocomposite for peripheral nerve regeneration.

Current commercially available nerve conduits fail to support nerve regeneration gaps larger than 30 mm in length due to the simple intra-luminal design of these conduits which are unable to biomimic the native neural environment. There is, therefore, a major clinical demand for new smart biomaterials, which can stimulate neuronal cell proliferation and migration, and facilitate nerve regeneration across these critical sized defects. In this study, we aimed to investigate Schwann cell (SC) behaviour seeded on the bioabsorbable version of the nanocomposite material, POSS modified poly (caprolactone) urea urethane (PCL), functionalised with arginine-glycine-aspartic acid (RGD) peptide. Successful synthesis of RGD peptide as well as the chemical structure of POSS-PCL nanocomposite film was investigated by Fourier transform infrared spectroscopy. Cell viability assay and morphological assessment were performed to investigate the cytocompatibility of the fabricated constructs. Successful immobilisation of RGD peptide onto the nanocomposite surface was confirmed by water contact angle, Brilliant Blue (BB) staining and thin layer chromatography. Both POSS-PCL and RGD-POSS-PCL nanocomposite scaffolds supported SC attachment, proliferation and morphological differentiation, important aspects for peripheral nerve regeneration. However, a significant increase in SC process length and morphological differentiation towards maturation was observed on the cells grown on RGD-POSS-PCL film. RGD-POSS-PCL nanocomposite demonstrated a significant improvement in SCs spreading and its integrin-dependent process outgrowth (P<0.05). Conduits made by POSS-nanocomposite may be suitable for the next generation of commercially available conduit required to meet current clinical demand in peripheral nerve regeneration and repair as they are currently undergoing in vivo preclinical study.

Effect of human urine on the tensile strength of sutures used for hypospadias surgery.

INTRODUCTION: Hypospadias is the most common congenital condition affecting between 1 in 250 and 300 live births. Even in experienced hands, surgery to repair this congenital defect can have a high complication rate. Wound dehiscence is reported to occur in 5% and fistula formation in 6%-40% depending on technique. The choice of suture material has been shown to affect the complication rate although there is (currently) no consensus about the best suture material to use. Ideally, the sutures used for urethroplasty should be absorbable while maintaining sufficient mechanical strength to support the wounds until they are self-supporting and able to resist urinary flow. Previous studies have compared the effects of human urine on different suture materials especially catgut. However, catgut is now banned in Europe. Our study examined the tensile and breaking strength as well as rate of degradation for four types of absorbable suture now commonly used for hypospadias repairs in the UK. MATERIAL AND METHODS: We examined the effect of prolonged storage (up to 27 days) in human urine on 6/0 gauge Vicryl, Vicryl Rapide, Monocryl and polydioxanone (PDS) sutures. These four suture materials are commonly used by the senior plastic consultant surgeon (NK) for hypospadias repairs. 50 mm sections of these suture materials were stored in either urine or saline as control. At specified time points, each suture was placed in a uniaxial load testing machine to assess the stress-strain profile and the mechanical load required to break the suture was measured. KEY RESULTS: Exposure to urine reduced the tensile and breaking strength of all the suture materials tested. PDS demonstrated the greatest resilience. Vicryl Rapide was the weakest suture and degraded completely by day 6. Vicryl and Monocryl had similar degradation profiles, but Vicryl retained more of its tensile strength for longer. CONCLUSIONS: There is a balance to be struck between the duration that a suture material must remain in any surgical wound and the risk that it causes foreign body effects. The results of this study suggest that Vicryl has the best characteristics for urethroplasty of the four suture materials tested.

Expression of mu opioid receptor splice variants mRNA in human blood lymphocytes: a peripheral marker for opioid addiction studies.

The expression of the human mu opioid receptor splice variants (hMOR-1A, hMOR-1O, hMOR-1X, and hMOR-1Y) mRNA in peripheral blood lymphocytes (PBLs) was explored in opioid addicted, methadone maintained, long-term abstinent, and control subjects. Real-time PCR showed that hMOR-1X and hMOR-1Y are not expressed in PBLs. The hMOR-1A mRNA was 0.33 fold down-regulated in abstinent and 1.94 fold up-regulated in methadone maintained subjects. The hMOR-1O mRNA was down-regulated 0.39 and 0.53 fold in abstinent and methadone maintained groups, respectively. Expression of both variants in addicted group was not different from controls. It is concluded that expression of hMOR-1A and hMOR-1O variants measured by a suggested peripheral marker can serve to identify people at risk for opioid addiction and also to evaluate the successfulness of methadone therapy.