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Correlation of donor age and telomerase activity with in vitro cell growth and replicative potential for dermal fibroblasts and keratinocytes

Ng MH, Aminuddin BS, Hamizah S, Lynette C, Mazlyzam AL, Ruszymah BH

J Tissue Viability, 2009 Nov;18(4):109-16.
PMID: 19632116 DOI: 10.1016/j.jtv.2009.06.003

Previous studies suggested telomerase activity as a determinant of cell replicative capacity by delaying cell senescence. This study aimed to evaluate the feasibility of adopting telomerase activity as a selection criterion for in vitro expanded skin cells before autologous transplantation. Fibroblasts and keratinoctyes were derived from the same consenting patients aged 9-69 years, and cultured separately in serum-supplemented and serum-free media, respectively. Telomerase activity of fresh and cultured cells were measured and correlated with cell growth rate, donor age and passage number. The results showed that telomerase activity and cell growth were independent of donor age for both cell types. Telomerase was expressed in freshly digested epidermis and dermis and continued expressing in vitro. Keratinocytes consistently showed 3-12 folds greater telomerase activity than fibroblast both in vivo and in vitro. Conversely, growth rate for fibroblast exceeded that of keratinocyte. Telomerase activity decreased markedly at Passage 6 for keratinocytes and ceased by Passage 3 for fibroblasts. The decrease or cessation of telomerase activity coincided with senescence for keratinocyte but not for fibroblast, implying a telomerase-regulated cell senescence for the former and hence a predictor of replicative capacity for this cell type. Relative telomerase activity for fibroblasts from the younger age group was significantly higher than that from the older age group; 69.7% higher for fresh isolates and 31.1% higher at P0 (p<0.05). No detectable telomerase activity was to be found at later subcultures for both age groups. Similarly for keratinocytes, telomerase activity in the younger age group was significantly higher (p<0.05) compared to that in the older age group; 507.7% at P0, 36.8% at P3 and the difference was no longer significant at P6. In conclusion, the study provided evidence that telomerase sustained the proliferation of keratinocytes but not fibroblasts. Telomerase activity is an important criterion for continued survival and replication of keratinocytes, hence its positive detection before transplantation is desirable. Inferring from our results, the use of keratinocytes from Passage 3 or lesser for construction of skin substitute or cell-based therapy is recommended owing to their sustained telomerase expression.
Trends of reactive hyperaemia responses to repetitive loading on skin tissue of rats - Implications for pressure ulcer prevention

Yapp JH, Kamil R, Rozi M, Mohtarrudin N, Loqman MY, Ezamin AR, et al.

J Tissue Viability, 2017 Aug;26(3):196-201.
PMID: 28438463 DOI: 10.1016/j.jtv.2017.03.002

Tissue recovery is important in preventing tissue deterioration, which is induced by pressure and may lead to pressure ulcers (PU). Reactive hyperaemia (RH) is an indicator used to identify people at risk of PU. In this study, the effect of different recovery times on RH trend is investigated during repetitive loading. Twenty-one male Sprague-Dawley rats (seven per group), with body weight of 385-485 g, were categorised into three groups and subjected to different recovery times with three repetitive loading cycles. The first, second, and third groups were subjected to short (3 min), moderate (10 min), and prolonged (40 min) recovery, respectively, while fixed loading time and pressure (10 min and 50 mmHg, respectively). Peak hyperaemia was measured in the three cycles to determine trends associated with different recovery times. Three RH trends (increasing, decreasing, and inconsistent) were observed. As the recovery time is increased (3 min vs. 10 min vs. 40 min), the number of samples with increasing RH trend decreases (57% vs. 29% vs. 14%) and the number of samples with inconsistent RH trend increases (29% vs. 57% vs. 72%). All groups consists of one sample with decreasing RH trend (14%). Results confirm that different recovery times affect the RH trend during repetitive loading. The RH trend may be used to determine the sufficient recovery time of an individual to avoid PU development.
Platelet-rich plasma with keratinocytes and fibroblasts enhance healing of full-thickness wounds

Law JX, Chowdhury SR, Saim AB, Idrus RBH

J Tissue Viability, 2017 Aug;26(3):208-215.
PMID: 28615133 DOI: 10.1016/j.jtv.2017.05.003

Advances in tissue engineering led to the development of various tissue-engineered skin substitutes (TESS) for the treatment of skin injuries. The majority of the autologous TESS required lengthy and costly cell expansion process to fabricate. In this study, we determine the possibility of using a low density of human skin cells suspended in platelet-rich plasma (PRP)-enriched medium to promote the healing of full-thickness skin wounds. To achieve this, full-thickness wounds of size 1.767 cm2 were created at the dorsum part of nude mice and treated with keratinocytes (2 × 104 cells/cm2) and fibroblasts (3 × 104 cells/cm2) suspended in 10% PRP-enriched medium. Wound examination was conducted weekly and the animals were euthanized after 2 weeks. Gross examination showed that re-epithelialization was fastest in the PRP+cells group at both day 7 and 14, followed by the PRP group and NT group receiving no treatment. Only the PRP+cells group achieved complete wound closure by 2 weeks. Epidermal layer was presence in the central region of the wound of the PRP+cells and PRP groups but absence in the NT group. Comparison between the PRP+cells and PRP groups showed that the PRP+cells-treated wound was more mature as indicated by the presence of thinner epidermis with single cell layer thick basal keratinocytes and less cellular dermis. In summary, the combination of low cell density and diluted PRP creates a synergistic effect which expedites the healing of full-thickness wounds. This combination has the potential to be developed as a rapid wound therapy via the direct application of freshly harvested skin cells in diluted PRP.
Stepease™ diabetic socks: An answer to efficacious indoor foot pressure relief- A prospective study

Soh EZF, Htwe O, Naicker AS, Nasirabadi AR, Ghazali MJ, Mohd Mustafah N, et al.

J Tissue Viability, 2020 May;29(2):104-109.
PMID: 32014382 DOI: 10.1016/j.jtv.2020.01.005

BACKGROUND: Diabetic foot ulcer is commonly seen in people with diabetes mellitus. Inadequate plantar pressure offloading has been identified as a contributing factor to development of diabetic foot ulcers. Various pressure off-loading footwear are widely available in the market but poor compliance has been reported especially for indoor usage. StepEase™ diabetic socks have been designed using Ethylene Vinyl Acetate (EVA) microspheres for better redistribution of plantar pressure. The objective of this study was to determine the efficacy of StepEase™ in redistributing the foot plantar pressure and to assess patients' satisfaction on the usage of the socks.
METHODS: This was a prospective non randomized clinical trial conducted on 31 patients with diabetes mellitus with high risk foot (King's classification stage II) over a 12 weeks period. Dynamic foot plantar pressure reading was recorded at day 0, 6 weeks and 12 weeks intervals, both barefoot and with StepEase™, using Novel Pedar-X system (Novel GmbH, Munich, Germany). Patients' satisfaction and usage practice were assessed by a questionnaire.
RESULTS: The mean age of subjects was 57.9 years with mean body mass index (BMI) of 26 kg/m2. The mean duration of diagnosis with diabetes mellitus was 10.2 years. The mean peak plantar pressure was found to be highest at the right forefoot and left heel region, 267.6 kPa (SD113.5 kPa) and 266.3 kPa (SD 94.6 kPa) respectively. There was a statistically significant reduction of mean peak pressure (P
Prevalence and prediction of pressure ulcers in admitted stroke patients in a tertiary care hospital

Farid J, Amin R, Sheikh MA, Irfan M, AlRuwaili R, Alruwaili M, et al.

J Tissue Viability, 2022 Nov;31(4):768-775.
PMID: 35941057 DOI: 10.1016/j.jtv.2022.07.010

Pressure ulcer (PU) is a localized injury to the skin or underlying tissues usually over a bony prominence, which results due to pressure or pressure in combination with shear. It is an expensive health care problem that have deterring impact on the length of hospitalization and cause extra nursing care time. Moreover, PUs negatively impacts patients' health related quality of life. High PUs prevalence figures were found in specialized hospital units such as intensive care unit (ICU), orthopedics, surgery, and also in stroke patients in medical units. The major purpose of this study is to assess the frequency of pressure ulcers in stroke patients at Ayub teaching hospital. The methodology used for carrying out the research was cross-sectional study conducted during months of September, October, and November 2020. Questionnaire was used to collect the data and well-informed written consent was taken from the patients. A total of 120 stroke patients were initially included with the intention to study the frequency of PUs among them. Different age groups were taken but majority (48.3%) belonged to the age group 31-60 years. Maximum patients were hypertensive (65%), while few of them were diabetic (35%). From the results of proposed work, it is found that out of 120 stroke patients, 75.8% presented with ischemic stroke while 24.2% presented with hemorrhagic stroke. 8.3% that is 10 out of 120 stroke patients developed pressure ulcers of grade 1 (1.7%), grade 2 (1.7%), grade 3 (2.5%), and grade 4 (2.5%) mostly in the sacral region (6.7%) and also on ankle (0.8%), and shoulder (0.8%) respectively. Patients in the study group had unsatisfactory hygiene (6.7%) were malnourished (11.7%) and were not using preventive mattresses (79.2%). Those at the risk of developing pressure ulcers were not being repositioned (6.7%) and did not had awareness (10%). Prevention and treatment used in ward is 100%. Conclusively, the frequency of pressure ulcers in stroke patients was determined to be 8.3% and the most frequent localization was sacrum. The PU care in this hospital is appropriate but still could be improved further by improving risk assessment, prevention specially use of air mattress and patient education regarding PUs. The main objective of the study is to identify the frequency of PUs in stroke patients and to highlight various factors that would avoid PUs development.
Cost-Effectiveness of PHMB & betaine wound bed preparation compared with standard care in venous leg ulcers: A cost-utility analysis in the United Kingdom

Cooper DM, Bojke C, Ghosh P

J Tissue Viability, 2023 Mar 16.
PMID: 36990897 DOI: 10.1016/j.jtv.2023.03.001

BACKGROUND: Wounds cost £8.3 billion per year in the United Kingdom (UK) annually. Venous leg ulcers (VLUs) account for 15% of wounds and can be complicated to heal, increasing nurse visits and resource costs. Recent wound bed preparation consensus recommends wound cleansing and biofilm disrupting agents. However, inert cleansers such as tap water or saline are inexpensive, an evaluation of evidence is required to justify the higher upfront costs of treatment with active cleansers. We undertook a cost-effectiveness analysis of the use of a biofilm disrupting and cleansing solution and gel, Prontosan® Solution and Gel X, (PSGX) (B Braun Medical), as compared to the standard practice of using saline solution, for treating VLUs.
METHODS: A Markov model was parameterised to one-year costs and health-related quality of life consequences of treating chronic VLUs with PSGX versus saline solution. Costs are viewed from a UK healthcare payer perspective, include routine care and management of complications. A systematic literature search was performed to inform the clinical parameters of the economic model. Deterministic univariate sensitivity analysis (DSA) and probabilistic sensitivity analysis (PSA) were undertaken.
RESULTS: For PSGX an Incremental Net Monetary Benefit (INMB) of £1,129.65 to £1,042.39 per patient (with a Maximum Willingness to Pay of £30k and £20k per QALY respectively), of which cost savings are £867.87 and 0.0087 quality-adjusted life years (QALYs) gain per patient. PSA indicates a 99.3% probability of PSGX being cost-effective over saline.
CONCLUSIONS: PSGX for the treatment of VLUs is dominant compared with saline solution in the UK with expected cost-savings within a year and improved patient outcomes.
Understanding the ideal wound healing mechanistic behavior using in silico modelling perspectives: A review

Abu Bakar N, Mydin RBSMN, Yusop N, Matmin J, Ghazalli NF

J Tissue Viability, 2024 Feb;33(1):104-115.
PMID: 38092620 DOI: 10.1016/j.jtv.2023.11.001

Complexity of the entire body precludes an accurate assessment of the specific contributions of tissues or cells during the healing process, which might be expensive and time consuming. Because of this, controlling the wound's size, depth, and dimensions may be challenging, and there is not yet an efficient and reliable chronic wound model representation. Furthermore, given the inherent challenges associated with conducting non-invasive in vivo investigations, it becomes peremptory to explore alternative methodologies for studying wound healing. In this context, biologically-realistic mathematical and computational models emerge as a valuable framework that can effectively address this need. Therefore, it might improve our approach to understanding the process at its core. This article will examines all facets of wound healing, including the kinds, pathways, and most current developments in wound treatment worldwide, particularly in silico modelling utilizing both mathematical and structure-based modelling techniques. It may be helpful to identify the crucial traits through the feedback loop of computer models and experimental investigations in order to build innovative therapies to cure wounds. Hence the effectiveness of personalised medicine and more targeted therapy in the healing of wounds may be enhanced by this interdisciplinary expertise.