Evaluation of the biocompatibility of fish skin collagen with the mesenchymal stem cells in in vitro cultures.

There are scarce published data suggesting, that collagen extracted from fish skin may be an attractive alternative to mammalian-derived collagen for the in vitro cell cultures. In this study, we investigated proliferation potential and differentiation capability into osteogenic and adipogenic lineages of rat adipose-derived mesenchymal stem cells (rASCs) and human adipose-derived mesenchymal stem cells (hASCs) cultured on collagen extracted from silver carp and African sharptooth catfish skins, compared with commercially available mammalian collagen and collagen-free culture dishes. Our results revealed no significant differences between fish collagen and mammalian collagen in supporting cell viability and proliferation capacity. Fish-derived collagen is a cheap material derived from production waste, does not contain transmissible pathogens of mammalian origin, supports human cell cultures at comparable level to conventional collagen sources, and may be considered as the product of choice for the in vitro cell cultures.

Effect of adipose-derived stem cells seeding and surgical prefabrication on composite scaffold vascularization.

The study aimed to evaluate an angiogenic effect of adipose-derived stem cells (ASCs) seeding and surgical prefabrication (placing a vascular pedicle inside the scaffold) on developed composite scaffolds made of poly-ε-caprolactone (PCL), ß-tricalcium phosphate (ß-TCP), and poly (lactic-co-glycolic acid) (PLGA) (PCL+ß-TCP+PLGA). Moreover, we aimed to compare our data with previously tested PCL scaffolds to assess whether the new material has better angiogenic properties. The study included 18 inbred male WAG rats. There were three scaffold groups (six animals each): with non-seeded PCL+ß-TCP+PLGA scaffolds, with PCL+ß-TCP+PLGA scaffolds seeded with ASCs and with PCL+ß-TCP+PLGA scaffolds seeded with ASCs and osteogenic-induced. Each rat was implanted with two scaffolds in the inguinal region (one prefabricated and one non-prefabricated). After 2 months from implantation, the scaffolds were explanted, and vessel density was determined by histopathological examination. Prefabricated ASC-seeded PCL+ß-TCP+PLGA scaffolds promoted greater vessel formation than non-seeded scaffolds (19.73 ± 5.46 vs 12.54 ± 0.81; p = .006) and those seeded with osteogenic-induced ASCs (19.73 ± 5.46 vs 11.87±2.21; p = .004). The developed composite scaffold promotes vessel formation more effectively than the previously described PCL scaffold.

The impact of electroconductive multifunctional composite nanofibrous scaffold on adipose-derived mesenchymal stem cells.

BACKGROUND: The development of tissue-engineered scaffolds with electrical properties is the primary motivation of novel regenerative medicine. Electroconductive scaffolds are designed to mimic the injured tissue environment's electrical properties and regulate cellular behavior - growth, proliferation, and differentiation - that could stimulate the injured nerve's regeneration. METHODS: We fabricated dedicated electroconductive scaffolds and customized an appropriate device with an external current supply to expose cells on the scaffold to electrical stimulation (ES). Next, we isolated rat adipose-derived stem cells (ASCs) and performed in vitro experiments that combine cells, an electroconductive scaffold, NGF (nerve growth factor), and ES (90 mV/mm, constant, for four days). Finally, we checked cellular activity as proliferation, viability, morphology, the neurogenic differentiation potential of ASCs, cell alignment, and karyotype. RESULTS: We observed that the electrical stimulation did not change the viability and chromosome stability of rat ASCs, but altered slightly proliferation compared to non-stimulated cells. The combined effect of a scaffold, NGF, and ES caused morphology changes and enhancement of ASCs neuronal differentiation as indicated in ßIII-tubulin expression, actin organization, and upregulation of neurogenic gene expression. CONCLUSIONS: We developed an electroconductive scaffold and customized device for in vitro study with many experimental variants. Based on our results, we presumed that the established study scheme - including an electroconductive scaffold, NGF and ES - is biocompatible and could guide ASCs to differentiate in neurogenic lineage, thus may be potentially applied in nerve injury regeneration.

Modified Histopathological Protocol for Poly-ɛ-Caprolactone Scaffolds Preserving Their Trabecular, Honeycomb-like Structure.

Poly-ɛ-caprolactone (PCL) is now widely studied in relation to the engineering of bone, cartilage, tendons, and other tissues. Standard histological protocols can destroy the carefully created trabecular and honeycomb-like architecture of PCL scaffolds, and could lead to scaffold fibers swelling, resulting in the displacement or compression of tissues inside the scaffold. The aim of this study was to modify a standard histopathological protocol for PCL scaffold preparation and evaluate it on porous cylindrical PCL scaffolds in a rat model. In 16 inbred Wag rats, 2 PCL scaffolds were implanted subcutaneously to both inguinal areas. Two months after implantation, harvested scaffolds were first subjected to µCT imaging, and then to histopathological analysis with standard (left inguinal area) and modified histopathological protocols (right inguinal area). To standardize the results, soft tissue percentages (STPs) were calculated on scaffold cross-sections obtained from both histopathological protocols and compared with corresponding µCT cross-sections. The modified protocol enabled the assessment of almost 10× more soft tissues on the scaffold cross-section than the standard procedure. Moreover, STP was only 1.5% lower than in the corresponding µCT cross-sections assessed before the histopathological procedure. The presented modification of the histopathological protocol is cheap, reproducible, and allows for a comprehensive evaluation of PCL scaffolds while maintaining their trabecular, honeycomb-like structure on cross-sections.

Long-term sensibility outcomes of secondary digital nerve reconstruction with sural nerve autografts: a retrospective study.

BACKGROUND: Recovery of sensibility after digital nerve injury is crucial for restoring normal hand function. We evaluated long-term outcomes of digital nerve reconstruction with autografts. METHODS: This retrospective study included patients who underwent secondary reconstruction of digital nerves with nerve autografting. Recovery of sensibility was evaluated based on the following: patient self-assessment, two-point discrimination (2PD), and a total sensation score (sum of proprioception, temperature sensation, and sharp/dull discrimination). Mixed models regression was used to study predictors of sensibility outcomes. The predictors analyzed were age, sex, smoking status, number of fingers involved in a patient (as a measure of injury severity), time to reconstruction, and time to follow-up. RESULTS: In 61 patients, 174 digital nerves in 126 fingers were reconstructed after an average of 33.1 weeks from injury. The mean follow-up was 6.4 years from reconstruction. The mean graft length was 3.6 cm. Self-rated sensibility in the affected area was very good in 13% of patients, good in 33%, satisfactory in 40%, and poor in 24%. 2PD at 6 mm was present in 17% of patients, at 10 mm in 12%, and at 15 mm in 18% (mean 2PD was 10.8). Proprioception was preserved in 107 (85%) fingers, sensation of temperature was preserved in 99 (75%) of fingers, and sharp/dull discrimination in 88 (70%) fingers. Time from injury to reconstruction was the only significant predictor of the total sensation score. CONCLUSION: Our data indicate that earlier reconstruction is associated with a favorable outcome.

Comparison of the Donor Age-Dependent and In Vitro Culture-Dependent Mesenchymal Stem Cell Aging in Rat Model.

Clinical experiments suggest that mesenchymal stem cells (MSCs) may be useful for tissue repair therapies or treatment of the autoimmune disorders. There is still lack of consensus concerning the age limit of MSC donors, majority of researchers suggest the autologous MSC therapies of patients not exceeding age limit of 55-60 yrs. The purpose of our study was to compare the selected parameters of MSCs from adipose tissue (adipose stem cell, ASC) collected from young and old rats of ages corresponding to patient's ages 25 yrs. and 80 yrs., respectively. The differences of parameters of ASCs from young and old animals were compared with the differences between ASCs from short-term (3 passage) and long-term (30 passage) in vitro culture. Cell morphology, surface marker expression, growth potential, metabolic activity, ß-galactosidase activity, clonogenic potential, angiogenic potential, and differentiation ability of ASCs from young and aged animals and from in vitro cultures at 3rd and 30th passages were compared and analyzed. It may be concluded that ASCs may be applied for autologous transplantations in aged patients. Comparison of ASC aging dynamics depending on host aging or in vitro culture duration suggests that long-term in vitro culture may affect ASCs more than natural aging process of their host. We suggest that ASCs expanded in vitro prior to their clinical use must be carefully screened for the possible aging effects resulting not only from donor age, but from the duration of their in vitro culture.

Bioactive Nanofiber-Based Conduits in a Peripheral Nerve Gap Management-An Animal Model Study.

The aim was to examine the efficiency of a scaffold made of poly (L-lactic acid)-co-poly(ϵ-caprolactone), collagen (COL), polyaniline (PANI), and enriched with adipose-derived stem cells (ASCs) as a nerve conduit in a rat model. P(LLA-CL)-COL-PANI scaffold was optimized and electrospun into a tubular-shaped structure. Adipose tissue from 10 Lewis rats was harvested for ASCs culture. A total of 28 inbred male Lewis rats underwent sciatic nerve transection and excision of a 10 mm nerve trunk fragment. In Group A, the nerve gap remained untouched; in Group B, an excised trunk was used as an autograft; in Group C, nerve stumps were secured with P(LLA-CL)-COL-PANI conduit; in Group D, P(LLA-CL)-COL-PANI conduit was enriched with ASCs. After 6 months of observation, rats were sacrificed. Gastrocnemius muscles and sciatic nerves were harvested for weight, histology analysis, and nerve fiber count analyses. Group A showed advanced atrophy of the muscle, and each intervention (B, C, D) prevented muscle mass decrease (p < 0.0001); however, ASCs addition decreased efficiency vs. autograft (p < 0.05). Nerve fiber count revealed a superior effect in the nerve fiber density observed in the groups with the use of conduit (D vs. B p < 0.0001, C vs. B p < 0.001). P(LLA-CL)-COL-PANI conduits with ASCs showed promising results in managing nerve gap by decreasing muscle atrophy.

Epidemiology of complex hand injuries treated in the Plastic Surgery Department of a tertiary referral hospital in Warsaw.

BACKGROUND: Trauma to the hand is common and potentially serious, impairing daily living and general quality of life. Patients are often unable to work for several months, with hand function improving only gradually. Here, we review the epidemiology of hand injuries treated at a tertiary referral hospital in Warsaw, Poland. MATERIAL AND METHODS: In this single-centre, retrospective study, we reviewed medical records of patients presenting to the A&E Unit of the Plastic Surgery Department, Centre of Postgraduate Medical Education in Warsaw, Poland, between January 2001 and December 2005. We assessed a number of patient and injury characteristics, including severity, scored with the Hand Injury Severity Scoring System (HISS), and time off work. RESULTS: Of 1091 patients with a hand injury, 84% were male and over half were under the age of 40 years. Hand injury commonly resulted in tendon damage (56.1%), especially to finger flexors (79%), and in skin loss (37.8%). Amputations occurred in 24.1% of cases, while fractures (9.6%) and nerve (6.1%) or joint (5.5%) damage were less common. HISS-graded injury severity was moderate in 28.6% of cases, over half of the patients suffered severe (25.5%) or major (26.5%) injuries, and minor injuries were relatively uncommon (19.4%). CONCLUSIONS: Amongst patients admitted to our Department, the most common injuries were tendon damage, skin loss, and amputations. Over half of the patients presented with severe or major injuries and took six months or longer to return to work, suggesting they were likely to face substantial social and economic consequences of their injury. LEVEL OF EVIDENCE: IV: retrospective series.

Scaffold vascularization method using an adipose-derived stem cell (ASC)-seeded scaffold prefabricated with a flow-through pedicle.

BACKGROUND: Vascularization is important for the clinical application of tissue engineered products. Both adipose-derived stem cells (ASCs) and surgical prefabrication can be used to induce angiogenesis in scaffolds. Our aim was to compare the angiogenic potential of ASC-seeded scaffolds combined with scaffold prefabrication with that of non-seeded, non-prefabricated scaffolds. METHODS: For prefabrication, functional blood vessels were introduced into the scaffold using a flow-through pedicle system. ASCs were isolated from rat fat deposits. Three-dimensional-printed cylindrical poly-ε-caprolactone scaffolds were fabricated by fused deposition modelling. Three groups, each containing six rats, were investigated by using non-seeded, ASC-seeded, and osteogenic induced ASC-seeded scaffolds. In each group, one rat was implanted with two scaffolds in the inguinal region. On the right side, a scaffold was implanted subcutaneously around the inferior epigastric vessels (classic prefabrication group). On the left side, the inferior epigastric vessels were placed inside the prefabricated scaffold in the flow-through pedicle system (flow-through prefabrication group). The vessel density and vascular architecture were examined histopathologically and by µCT imaging, respectively, at 2 months after implantation. RESULTS: The mean vessel densities were 10- and 5-fold higher in the ASC-seeded and osteogenic induced ASC-seeded scaffolds with flow-through prefabrication, respectively, than in the non-seeded classic prefabricated group (p < 0.001). µCT imaging revealed functional vessels within the scaffold. CONCLUSION: ASC-seeded scaffolds with prefabrication showed significantly improved scaffold vasculogenesis and could be useful for application to tissue engineering products in the clinical settings.

Comparison of adipose stem cells sources from various locations of rat body for their application for seeding on polymer scaffolds.

Adipose tissue yields adult adipose stem cells (ASCs) in large quantities via less-invasive methods. These cells are of interest owing to their modulating properties and paracrine activities, which can be harnessed in regenerative medicine. Many studies on the use of rat fat tissue in an autologous animal model have been conducted; however, the different locations to obtain stromal vascular fraction of rat fat depots have not been fully characterized. The purpose of the current study was to identify optimal source of ASC from various locations of rat body. Animal experiments in vitro revealed that fat depots from cervical fat are an optimal ASC source. A high ASC yield facilitates subsequent studies on autologous transplantation in rats. The secondary objective was to compare the efficiency of osteoinductive media composition and evaluate of osteogenic potential of ASCs for seeding on scaffolds for bone repair. Scaffolds were assessed in vitro, using rat adipose stem cells and three-dimensional (3D) scaffolds comprising polycaprolactone (PCL) or polycaprolactone covered with tricalcium phosphate (PCL + 5%TCP). Seeded ASCs adhere to the surface and migrate to the scaffolds. Upon staining and determining alkaline phosphatase levels, PCL + 5%TCP scaffolds performed better than PCL scaffolds. Furthermore, growth factors such as BMP2 and FGF2 significantly increased ASC mineralization and induced osteogenesis (p < 0.05). Our results may help select and develop pre-clinical animal model for confirming the use of ASC, alone or in association with appropriate biomaterials for bone repair.

Aspects of Modern Biobank Activity - Comprehensive Review.

Biobanks play an increasing role in contemporary research projects. These units meet all requirements to regard them as a one of the most innovative and up-to-date in the field of biomedical research. They enable conducting wide-scale research by the professional collection of biological specimens and correlated clinical data. Pathology units may be perceived roots of biobanking. The review aims at describing the concept of biobanks, their model of function and scientific potential. It comprises the division of biobanks, sample preservation methods and IT solutions as well as guidelines and recommendations for management of a vast number of biological samples and clinical data. Therefore, appropriate standard operating procedures and protocols are outlined. Constant individualization of diagnostic process and treatment procedures creates the niche for translational units. Thus, the role of biobanks in personalized medicine was also specified. The exceptionality of biobanks poses some new ethical-legal issues which have various solutions, in each legal system, amongst the world. Finally, distribution and activity of European biobanks are mentioned.

A novel computer system for the evaluation of nasolabial morphology, symmetry and aesthetics after cleft lip and palate treatment. Part 1: General concept and validation.

PURPOSE: The need for a widely accepted method suitable for a multicentre quantitative evaluation of facial aesthetics after surgical treatment of cleft lip and palate (CLP) has been emphasized for years. The aim of this study was to validate a novel computer system 'Analyse It Doc' (A.I.D.) as a tool for objective anthropometric analysis of the nasolabial region. MATERIALS AND METHODS: An indirect anthropometric analysis of facial photographs was conducted with the A.I.D. system and Adobe Photoshop/ImageJ software. Intra-rater and inter-rater reliability and the time required for the analysis were estimated separately for each method and compared. RESULTS: Analysis with A.I.D. system was nearly 10-fold faster than that with the reference evaluation method. The A.I.D. system provided strong inter-rater and intra-rater correlations for linear, angular and area measurements of the nasolabial region, as well as a significantly higher accuracy and reproducibility of angular measurements in submental view. No statistically significant inter-method differences were found for other measurements. CONCLUSIONS: The hereby presented novel computer system is suitable for simple, time-efficient and reliable multicenter photogrammetric analyses of the nasolabial region in CLP patients and healthy subjects.

Incomplete excision of basal cell carcinoma (BCC) in the head and neck region: to wait, or not to wait?

INTRODUCTION: Recurrence rates for incompletely excised basal cell carcinoma (BCC) vary widely in the literature. Clinical observation is a commonly accepted method of follow up, however such management of these lesions still remains controversial. AIM: To evaluate the rate and factors associated with the recurrence of BCC of the head and neck region after incomplete excision. MATERIAL AND METHODS: Medical records of 135 patients with 156 incompletely excised BCCs of the head and neck region were analyzed retrospectively. The primary outcome was the rate of recurrence. Additionally, a correlation of recurrence to clinical and morphological factors was analyzed. RESULTS: Recurrence occurred in 72 (46%) lesions. The mean interval to recurrence was 20 months. In each category of factors, the highest relative risk of recurrence was correlated to: location on the scalp - 2.27, diameter over 2 cm - 1.21, nodular clinical form - 1.29, morpheaform histopathological type - 1.67, recurrent lesion - 1.88, irradicality of excision in the lateral margin - 1.24 and closure of the skin defect with the split-thickness skin graft - 1.42 relative risk. CONCLUSIONS: Observation is an acceptable management option as less than a half of incompletely excised BCCs recurred and needed further treatment. As 85% of recurrences occur within 3 years after operation, clinical observation should be particularly careful during this period, however long-term recurrence should not be underestimated.

[Mesenchymal stem cells]. / Mezenchymalne komórki macierzyste.

The multipotential progenitor cells called ,Mesenchymal Stem Cells" (MSC) are capable of differrentiation at least into bone, cartilage, and adipose tissues. The commonly recognized role of these cells is the formation of connective tissue which participates in formation of every organ. The progeny of MSC produces also the hematopoietic microenvironment, recently it have been documented that these cells are capable of the modulation of the immune system activities. MSC are isolated from the tissues of fetal origin (umbilical cord, cord blood, or placenta), or from several adult donor sites, in particular from bone marrow and adipose tissue which are most useful for practical purposes. The capability of multipotential differentiation, immunomodulation, and the regulation of the endogenous tissue repair are the reasons why mesenchymal stem cells are widely applied for regenerative medicine purposes.

[Correction of secondary bilateral cleft lip deformity - outcome evaluation]. / Korekcja znieksztalcenia wargi po zeszyciu obustronnego rozszczepu wargi, wyrostka zebodolowego i podniebienia - ocena wyników leczenia.

UNLABELLED: One of the methods for correction of "whistle" deformity following a primary bilateral cleft lip repair is a Skoog procedure. It consists of orbicularis oris muscle mobilization and suture following vermilion closure in a V-Y manner. The aim of the study was to evaluate late results of Skoog procedure in "whistle" deformity correction. MATERIAL AND METHODS: Between 2000 and 2009 12 patients with whistle deformity underwent Skoog procedure. All of them were operated by the same surgeon. Aesthetic outcome in terms of lip fullness, symmetry and overall appearance was evaluated by patient and three independent examiners in four-point scale (1 - dissatisfied, 4 - very good). Pre- and postoperative vertical vermilion height was calculated bydigital analysis of photographs. in addition, complications and patient problems such as lip pain, microstomia, smiling or speech disturbances and scabbing of malpositioned wet vermilion were recorded. RESULTS: Six patients (4 men, 2 women, aged 23-44 years, mean 29 years) were included into the study. The average follow-up was 69.5 months. Patients higher evaluated overall appearance (mean 3.83) than symmetry (mean 3.16) and lip fullness (mean 3.16). Lip symmetry evaluated by independent examiners was assessed higher (mean 3.5) than overall appearance (mean 3.33) and lip fullness (mean 3). Digital analysis of photographs shows that mean vertical vermilion height increased after Skoog procedure from 34 to 142% (mean 71.5%). Besides acceptable scabbing of malpositioned wet vermilion no complications and patient problems occurred. CONCLUSIONS: Skoog procedure provided a simple and reliable option for the correction of "whistle" deformity.

[Frontalis suspension using autogenous fascia lata--evaluation of long-term outcome]. / Podwieszenie powiek górnych na paskach powiezi szerokiej uda--ocena póznych wyników.

PURPOSE: To evaluate long-term outcome of frontalis suspension using autogenous fascia lata (frontalis suspension - FS) which is indicated in the treatment of severe blepharoptosis with minimal levator function. MATERIAL AND METHODS: Fourty one patients (23 females, 18 males, aged 6-79 years, mean 41.4 years) who underwent FS between 1999 and 2009 were evaluated. Mean follow-up was 55 months. Functional outcome was measured on digital photographs by analysis of an upper eyelid margin position below the superior limbus. Following criteria were used: very good (< 3 mm), good (3-5 mm), unsatisfactory (> 5 mm). Aesthetic outcome was assessed in terms of lid contour, symmetry of lid height and lid crease. Patient satisfaction and life quality improvement were evaluated in the survey. RESULTS: Functional outcome was very good in 36%, good in 59.3%, unsatisfactory in 4.7%. Recurrence occured in 4.6% and complications in 10.9% (lagophthalmos--6.2%, entropion--4.7%). Very good aesthetic outcome regarding lid contour, symmetry of lid height and lid crease was achieved in 76.6%, 53.6% and 51.6%, respectively. Patient satisfaction and life quality improvement was also very high. There were only 1.6% unsatisfied patients. CONCLUSIONS: FS is an efficient method in treatment of severe blepharoptosis.

[Gorlin-Goltz syndrome--a case report]. / Zespól Gorlina-Goltza--opis przypadku.

UNLABELLED: The Gorlin-Goltz syndrome (GGS) (the nevoid basal cell carcinoma syndrome-NBCCS) is an autosomal dominant syndrome caused by mutations found on chromosome 9. The syndrome is characterized by increased predisposition to develop a basal cell carcinoma and associated with multiorgan anomalies. THE AIM OF THIS WORK: To present a case of GGS and explain modern standards of care for patients with this syndrome. CASE REPORT: Authors report the case of a 36-year-old patient who was admitted to the Plastic Surgery Clinic due to numerous basal cell carcinomas. Previously patient underwent an orthopaedic, neurologic, dermatologic, stomatologic and surgery treatment due to particular anomalies which characterize this syndrome. Comprehensive interview and broadening of the diagnostics enabled to diagnose GGS and to introduce the appropriate treatment. CONCLUSIONS: GGS is a multidisciplinary problem and widespread knowledge of this syndrome could accelerate the diagnosis process. Early diagnosis of GGS allows to introduce the secondary prophylaxis and to apply the appropriate treatment to slow the progress of the syndrome.