Toxicity study in a pig model of intraperitoneal collagenase as an "enzymatic scalpel" directed to break stroma in order to generate a new perspective for peritoneal carcinomatosis approach: an experimental research.

BACKGROUND: This study aimed to measure the toxicity resulting from collagenase administration to the peritoneal cavity in a pig model as a preliminary step to break down the stroma surrounding tumors. METHODS: Eight pigs were treated with 2 different collagenase concentrations previously tested in rats by our group. Time and temperature were controlled using a peritoneal lavage system (PRS System, Combat Medical Ltd.) identical to that used in human surgeries through hyperthermic intraperitoneal chemotherapy (HIPEC); 2 additional pigs were treated with peritoneal lavage only. Samples of blood and peritoneal fluid were collected pre-treatment, immediately after treatment, and 24 h postoperatively. In addition, histological studies and blood collagenase levels were measured. RESULTS: No complications were observed during the surgeries. Intraoperative images evidenced the release of peritoneal tissue during collagenase treatment. After surgery, the animals showed no signs of pain. Diet and mobility were normal at 4 h postoperatively, and there were no significant differences in hematologic or biochemical parameters. Quantification of MMP1 and MMP2 in all samples as measured by absorbance showed no differences in blood collagenase levels between pre-treatment, post-treatment, and 24 h postoperatively. None of the animals treated with collagenase showed peritoneal adhesions during the second surgery. Histologically, peritoneal organs and serous structures did not show any microscopic alterations associated with collagenase treatment in any group. CONCLUSION: Lavage of the peritoneal cavity with doses of up to 100,000 collagen digestion units/animal for 30 min is safe and removes connective tissue from the peritoneal cavity.

Wide-awake anesthesia in Dupuytren's contracture treated with collagenase.

The injection of collagenase followed by cord manipulation is one of the most popular treatments for Dupuytren's contracture. This is traditionally performed under local anesthesia or regional nerve block potentially with sedation. Neither the treatment with collagenase, nor the wide-awake anesthesia are novel techniques for hand surgeons. Nevertheless, we report the first experience of cord manipulation using the wide-awake approach. In this prospective study, we compared the pain perception of patients who underwent wide-awake anesthesia versus traditional local anesthesia. We recorded the pain sensation on a visual analog scale (VAS) (0 to 10) during anesthetic injection, during cord manipulation and before discharge. Wide-awake anesthesia significantly reduced pain levels during anesthetic injection (p=0.003) and cord manipulation (p=0.0009). Pain levels did not differ significantly right before discharge in the two groups (p=0.54). Wide-awake anesthesia can be successfully applied to cord manipulation after collagenase injection in Dupuytren's contracture. This way, it is possible to improve the patient's subjective perspective of the procedure.

Clinical Value of Debriding Enzymes as an Adjunct to Standard Early Surgical Excision in Human Burns: A Systematic Review.

Burns are a significant public health burden worldwide. In addition to those who die, millions remain with life-threatening deformities and disabilities resulting in stigma and rejection. Surgical excision is currently the standard of care for removing necrotic tissues in burn wounds to prepare the wound bed for grafting or enhancing the healing process. However, there is a growing interest on enzymatic debridement as an adjunct therapy in burn wounds. The aim of this study was to investigate clinical trials using debriding agents for burn wound in humans in a systematic review. This was a systematic review of electronic databases including CINAHL, PubMed, Ovid Medline, Web of Science, Google Scholar, and Embase from January 1969 to February 2019. The study protocol was registered in PROSPERO registry. The following keywords were searched: "burn wounds", "enzymatic debridement", "papain", "papain-urea", "pine apple", "Bromelain", "collagenases", "Nexobrid", "Debrase", "Debridase", "Actinidia deliciosa", "Sutilains", "Debrace", "piruvat acid". Those studies fulfilling the inclusion and exclusion criteria with low score of bias based on Cochrane Bias Tool were reviewed. Sixteen investigations fulfilled our inclusion criteria to be reviewed. Six, seven, and three clinical trials on humans were found regarding collagenase, bromelain, and miscellaneous agents. Collagenase has been reported to be effective in burns below 25% of TBSA, especially in outpatients' clinics. However, Nexobrid has been shown to be effective in deep burns and decreases the percentage of graft without significant adverse effects. There was not enough evidence supporting the clinical values of Papain, Sutilains, Urea, etc. Surgical excision still remains the standard of care for burn wounds debridement. However, enzymatic debridement, especially Bromelain might help to reduce sessions for surgical debridement or area under graft as an adjunct treatment. Despite the fact, more studies with larger sample sizes and with less conflicts of interest are needed to clearly elucidate the exact role of Bromelain.

Time to start putting down the knife: A systematic review of burns excision tools of randomised and non-randomised trials.

AIMS: Dermal preservation during acute burn excision is key to obtaining superior healing/scar outcomes, however, determining the most appropriate excision tool is an ongoing challenge. Novel tool development means the knife is no longer our only option, yet for the majority it remains the gold standard. This systematic review aims to evaluate evidence for burns excision approaches (knife/hydrosurgery/enzymatic). METHODS: CENTRAL, EMBASE, MEDLINE (1946-2017) were searched with MeSH terms: 'debridement', 'burns', 'sharp', 'enzymatic', 'hydrosurgery'. Relevant randomised control trials (RCTs)/non-randomised controlled case series/trials were extracted/analysed. In vitro/burn non-specific studies were excluded. Main methodological parameters were intervention/excision efficacy. RESULTS: Eighteen articles met inclusion criteria (n=7148): three were RCTs, involving comparator enzymatic (NexoBrid™ (EDNX)) or hydrosurgical (Versajet™) excision to surgical Standard of Care. Both showed statistically significant decreased need for excisional excision and auto-grafting by viable tissue preservation allowing spontaneous healing by epithelialisation. CONCLUSION: Level 1 Evidence comparing excision modalities for acute burns is sparse. Although early excision with a knife is still often considered best practice, there is no tool choice consensus or robust comparison with alternate, possibly superior, tools. EDNX or Versajet™ should be considered alternatively. Further RCTs are indicated, with regards final scar outcomes and to allow consensus within current evidence.

Dressings and topical agents for treating pressure ulcers.

BACKGROUND: Pressure ulcers, also known as bedsores, decubitus ulcers and pressure injuries, are localised areas of injury to the skin or the underlying tissue, or both. Dressings are widely used to treat pressure ulcers and promote healing, and there are many options to choose from including alginate, hydrocolloid and protease-modulating dressings. Topical agents have also been used as alternatives to dressings in order to promote healing.A clear and current overview of all the evidence is required to facilitate decision-making regarding the use of dressings or topical agents for the treatment of pressure ulcers. Such a review would ideally help people with pressure ulcers and health professionals assess the best treatment options. This review is a network meta-analysis (NMA) which assesses the probability of complete ulcer healing associated with alternative dressings and topical agents. OBJECTIVES: To assess the effects of dressings and topical agents for healing pressure ulcers in any care setting. We aimed to examine this evidence base as a whole, determining probabilities that each treatment is the best, with full assessment of uncertainty and evidence quality. SEARCH METHODS: In July 2016 we searched the Cochrane Wounds Specialised Register; the Cochrane Central Register of Controlled Trials (CENTRAL); Ovid MEDLINE; Ovid MEDLINE (In-Process & Other Non-Indexed Citations); Ovid Embase and EBSCO CINAHL Plus. We also searched clinical trials registries for ongoing and unpublished studies, and scanned reference lists of relevant included studies as well as reviews, meta-analyses, guidelines and health technology reports to identify additional studies. There were no restrictions with respect to language, date of publication or study setting. SELECTION CRITERIA: Published or unpublished randomised controlled trials (RCTs) comparing the effects of at least one of the following interventions with any other intervention in the treatment of pressure ulcers (Stage 2 or above): any dressing, or any topical agent applied directly to an open pressure ulcer and left in situ. We excluded from this review dressings attached to external devices such as negative pressure wound therapies, skin grafts, growth factor treatments, platelet gels and larval therapy. DATA COLLECTION AND ANALYSIS: Two review authors independently performed study selection, risk of bias assessment and data extraction. We conducted network meta-analysis using frequentist mega-regression methods for the efficacy outcome, probability of complete healing. We modelled the relative effectiveness of any two treatments as a function of each treatment relative to the reference treatment (saline gauze). We assumed that treatment effects were similar within dressings classes (e.g. hydrocolloid, foam). We present estimates of effect with their 95% confidence intervals for individual treatments compared with every other, and we report ranking probabilities for each intervention (probability of being the best, second best, etc treatment). We assessed the certainty (quality) of the body of evidence using GRADE for each network comparison and for the network as whole. MAIN RESULTS: We included 51 studies (2947 participants) in this review and carried out NMA in a network of linked interventions for the sole outcome of probability of complete healing. The network included 21 different interventions (13 dressings, 6 topical agents and 2 supplementary linking interventions) and was informed by 39 studies in 2127 participants, of whom 783 had completely healed wounds.We judged the network to be sparse: overall, there were relatively few participants, with few events, both for the number of interventions and the number of mixed treatment contrasts; most studies were small or very small. The consequence of this sparseness is high imprecision in the evidence, and this, coupled with the (mainly) high risk of bias in the studies informing the network, means that we judged the vast majority of the evidence to be of low or very low certainty. We have no confidence in the findings regarding the rank order of interventions in this review (very low-certainty evidence), but we report here a summary of results for some comparisons of interventions compared with saline gauze. We present here only the findings from evidence which we did not consider to be very low certainty, but these reported results should still be interpreted in the context of the very low certainty of the network as a whole.It is not clear whether regimens involving protease-modulating dressings increase the probability of pressure ulcer healing compared with saline gauze (risk ratio (RR) 1.65, 95% confidence interval (CI) 0.92 to 2.94) (moderate-certainty evidence: low risk of bias, downgraded for imprecision). This risk ratio of 1.65 corresponds to an absolute difference of 102 more people healed with protease modulating dressings per 1000 people treated than with saline gauze alone (95% CI 13 fewer to 302 more). It is unclear whether the following interventions increase the probability of healing compared with saline gauze (low-certainty evidence): collagenase ointment (RR 2.12, 95% CI 1.06 to 4.22); foam dressings (RR 1.52, 95% CI 1.03 to 2.26); basic wound contact dressings (RR 1.30, 95% CI 0.65 to 2.58) and polyvinylpyrrolidone plus zinc oxide (RR 1.31, 95% CI 0.37 to 4.62); the latter two interventions both had confidence intervals consistent with both a clinically important benefit and a clinically important harm, and the former two interventions each had high risk of bias as well as imprecision. AUTHORS' CONCLUSIONS: A network meta-analysis (NMA) of data from 39 studies (evaluating 21 dressings and topical agents for pressure ulcers) is sparse and the evidence is of low or very low certainty (due mainly to risk of bias and imprecision). Consequently we are unable to determine which dressings or topical agents are the most likely to heal pressure ulcers, and it is generally unclear whether the treatments examined are more effective than saline gauze.More research is needed to determine whether particular dressings or topical agents improve the probability of healing of pressure ulcers. The NMA is uninformative regarding which interventions might best be included in a large trial, and it may be that research is directed towards prevention, leaving clinicians to decide which treatment to use on the basis of wound symptoms, clinical experience, patient preference and cost.

Dermoabrasión como alternativa de tratamiento en quemaduras tipo AB en pacientes pediátricos. Experiencia del Servicio de Cirugía Plástica, Unidad de Quemados del Hospital Pediátrico Fernando Bareyro / Dermoabrasion for treatment in pediatric burns

La dermoabrasión en combinación con sulfadiazina de plata y lidocaína, como tratamiento para las quemaduras AB en pacientes pediátricos, presenta ventajas en cuanto a los resultados que ofrece, ya que es un método sencillo y reproducible. Como desventaja presenta que se requieren varias sesiones de tratamiento y en algunos casos, donde se produjo profundización de la lesión, se requirió cobertura con injerto de pie

The vegetal biomembrane in the healing of chronic venous ulcers.

BACKGROUND: The vegetal biomembrane has been used to treat cutaneous ulcers. OBJECTIVES: To assess the role of the vegetal biomembrane on the chronic venous ulcers treatment compared to treatment with collagenase cream. METHODS: Fourteen patients were selected to be treated with vegetal biomembrane and 7 with Fibrase(®) (CONTROL), followed clinically and photographically by the Wound Healing Index by ImageJ during 120 days and biopsied on the 1(st) and 30(th) days for histological examination. RESULTS: The vegetal biomembrane was better in promoting healing of the ulcers, especially on the inflammatory phase, confirmed by abundant exudation and wound debridement than the CONTROL group, on the 30th day. There was a greater tendency to angiogenesis followed by re-epithelialization with highest wound healing index on the 90(th) and 120(th) days. CONCLUSION: A combined analysis of clinical and histopathological findings suggests that the vegetal biomembrane acted as a factor inducing wound healing, especially on the inflammatory phase, confirmed by abundant exudation of the lesions promoting the transformation of the microenvironment of the chronic venous ulcers, and also stimulating angiogenesis and subsequent re-epithelialization.

The vegetal biomembrane in the healing of chronic venous ulcers / Biomembrana vegetal na cicatrização de úlceras venosas crônicas

BACKGROUND: The vegetal biomembrane has been used to treat cutaneous ulcers. OBJECTIVES: To assess the role of the vegetal biomembrane on the chronic venous ulcers treatment compared to treatment with collagenase cream. METHODS: Fourteen patients were selected to be treated with vegetal biomembrane and 7 with Fibrase®(CONTROL), followed clinically and photographically by the Wound Healing Index by ImageJ during 120 days and biopsied on the 1st and 30th days for histological examination. RESULTS: The vegetal biomembrane was better in promoting healing of the ulcers, especially on the inflammatory phase, confirmed by abundant exudation and wound debridement than the CONTROL group, on the 30th day. There was a greater tendency to angiogenesis followed by re-epithelialization with highest wound healing index on the 90th and 120th days. CONCLUSION: A combined analysis of clinical and histopathological findings suggests that the vegetal biomembrane acted as a factor inducing wound healing, especially on the inflammatory phase, confirmed by abundant exudation of the lesions promoting the transformation of the microenvironment of the chronic venous ulcers, and also stimulating angiogenesis and subsequent re-epithelialization.

FUNDAMENTOS: A biomembrana vegetal tem sido usada para tratamento de úlceras cutâneas. OBJETIVOS: Avaliar a ação da biomembrana vegetal no tratamento de úlceras venosas crônicas, comparando-a ao tratamento à base de colagenase. MÉTODOS: Foram selecionados 14 pacientes tratados com biomembrana vegetal e sete com Fibrase® (grupo controle), acompanhados clínico-fotograficamente pelo índice de cicatrização das úlceras (ICU) por 120 dias, por meio do software ImageJ, e biopsiados no primeiro e 30º dias para estudo histopatológico. RESULTADOS: A biomembrana vegetal foi superior em relação ao controle na cicatrização das úlceras no 30º dia, especialmente na fase inflamatória, confirmada pela exsudação abundante e pelo desbridamento. Houve tendência superior à angiogênese seguida de reepitelização com maiores ICUs no 90º e 120º dias. CONCLUSÃO: A análise conjunta dos achados clínicos e histopatológicos sugere que a biomembrana vegetal atuou como um fator indutor da cicatrização, especialmente na fase inflamatória, confirmada pela exsudação abundante das lesões, promovendo a transformação do microambiente das úlceras venosas crônicas e estimulando a angiogênese e a posterior reepitelização.

[Chemonucleolysis and intradiscal electrothermal therapy: what is the current evidence?]. / Chemonukleolyse und intradiskale Elektrotherapie: was ist die gegenwärtige Evidenz?

We evaluated the efficacy and safety of chemonucleolysis and intradiscal electrothermal therapy (IDET) on the basis of the data presented in recently published papers with respect to pain relief, function, and complication rates. Detailed searches for English and German articles published between 2003 and 2008 were performed in a number of electronic databases. Further publications were identified by manual search. For summarizing the evidence, we considered only systematic reviews and controlled studies. The internal validity of reviews and studies was judged by two authors independently. Data extraction was performed by one author, and the extracted data was checked for completeness and correctness by a second author. The evidence of the efficacy of chemonucleolysis using chymopapain or collagenase is summarized in two recent, high-quality systematic reviews. We found 5 controlled studies evaluating nucleolysis using an oxygen-ozone mixture (O (2)O (3)-nucleolysis). Some of those studies were of limited methodological quality, but all showed the efficacy of O (2)O (3)-nucleolysis in comparison to microdiscectomy or the use of alternative substances. There is hardly any data regarding O (2)O (3)-nucleolysis complications. Regarding IDET, the authors of the 6 identified systematic reviews come to different conclusions about the efficacy of the procedure. The results of the 3 included controlled IDET studies, of which 2 are of high methodological quality, are also conflicting. The complication rates range from 0 to 15 %. In summary, the evidence of efficacy is presently more compelling for chemonucleolysis than for IDET. This may also be because indications for chemonucleolysis are more firmly established. However, safety aspects should be better evaluated and presented in the literature.

Non-surgical therapy of Peyronie's disease.

The present paper provides a review of the available non-surgical treatments for Peyronie's disease (PD). A review of published literature on oral, intralesional, external energy and iontophoresis therapies for PD was performed, and the published results of available treatment options reviewed. The authors recommendations for appropriate non-surgical management of PD are provided. Although there are many published reports that show the efficacy of non-surgical therapies for PD, there is a lack of large scale, multicenter controlled clinical trials, which makes treatment recommendations difficult. Careful review of the literature does suggest that there are treatment options that make scientific sense and appear to stabilize the disease process, reduce deformity, and improve function. Offering no treatment at all will encourage our patients to pursue alternative treatments, which might do harm, and misses the opportunity to do some good. Clearly further work is necessary to develop safe and effective non-surgical treatments for PD.

Dynamics of hydration of cicatricial tissue during local therapy with collagenase preparations.

The components of complex dielectric permeability of cicatricial tissue at the frequency of 55 GHz were studied during local therapy of cicatrices with Fermenkol (a complex of collagenolytic proteases). Electrical characteristics of tissues in the microwave range were interpreted in terms of hydration parameters (total content of water in tissues and the ratio of structured and free water). The rightfulness of interpretations is discussed on the basis of the results of measurements of cell suspension models (native blood and its fractions brought to a standard hematocrit). The results attest to a relationship between hydration of the cicatricial tissue and its morphology and function.

Discernment of adipose versus nervous tissue: a novel adjunct solution in lipomyelomeningocele surgery.

OBJECTIVE: To determine a solution capable of discerning adipose versus nervous tissue, to aid in surgical separation of the adipose tissue which appears to be visually indistinguishable from nervous tissue in lipomyelomeningoceles (LMMs). METHODS: The following solutes (in normal saline) were investigated, both at 25 and 37 degrees C: beta-carotene, vitamin D, vitamin E, lecithin, hydrogen peroxide, lipase, protease, hyaluronidase, partially purified collagenase, purified collagenase, trypsin, trypsin plus purified collagenase and non-solute-containing saline (control). Each solution was applied to a pediatric lipoma to determine gross effects over a period of approximately 30 min. If a solution appeared to affect the adipose tissue grossly, studies of functional in vivo sensory evoked and spontaneous potentials using that particular solution were conducted upon sheep spinal cord, nerve roots, dura and peripheral nerve. Additionally, histological studies were conducted to determine the effect of that solution upon adipose tissue, spinal cord, myelin, dura and nerve roots. RESULTS: Of all solutions investigated, partially purified collagenase type 1 (T1C; Lot M0M4322, Code CLS-1, Worthington Biochemical Corporation, Lakewood, N.J., USA) at 37 degrees C was the most successful in grossly altering the consistency and appearance of adipose tissue. This change was more apparent over 20-30 min following application of the solution to the adipose tissue. Solutions not containing T1C did not show appreciable results; purified collagenase plus trypsin did not appear comparable or superior to T1C. No significant histological or functional change was noted when comparing the spinal cord, nerve rootlets, myelin, dura or peripheral nerve from the T1C-treated group versus normal (untreated) control groups. CONCLUSION: T1C appears to be a potentially effective solution for application during LMM surgery in the acute setting, and such use of an adjunct solution may significantly aid in the safe surgical resection of LMMs. Pending further research, this technique may be applied for other indications which require discernment or alteration of adipose versus nervous tissue.

[Effect of improved topical agents on healing time of deep second-degree burn wound].

OBJECTIVE: With the recognization of the mechanism of wound healing, some topical agents are created and applied in trauma to improve the healing rate of wounds. The main purpose of this study is to investigate the effect of some topical agents on the healing rate of deep second-degree burn wounds. METHODS: One thousand five hundred and sixty-three patients with deep second-degree burn wounds(total burn surface area < or = 10%) were involved in this study from January 1982 to December 1999. According to the application time of different treating measures including supplement of Zn, application of growth factors and collagenase, the patients were divided into 3 groups, wound healing rates were compared. RESULTS: Before 1991, none of special topical agents were used, and the healing time of deep second-degree burn wounds was(23.8 +/- 3.5) days. From 1991 to 1996, with the topical application of SD-Ag-Zn, which can provide Zn for cells taking part in wound healing, the healing time of deep second-degree burn wounds was (20.6 +/- 3.2) days, earlier than no special topical agents (P < 0.05). From 1997 to 1999, growth factors such as basic fibroblast growth factor(bFGF) and epithelial growth factor (EGF) and collagenases were applied in wound treatment combining with SD-Ag-Zn, wound healing time was (16.2 +/- 2.8) days, earlier than no special topical agents (P < 0.01) and simple SD-Ag-Zn application (P < 0.05). CONCLUSION: It indicates that the improvement of topical agents can accelerate wound healing speed.

Crab collagenase in wound debridement.

A comparative study was performed of the caseinolytic, necrolytic and fibrinolytic activities of a crab collagenase product prepared from the hepatopancreas of the king crab (Paralithodes camtschatica) and four enzyme preparations (trypsin/chymotrypsin and protease complexes isolated from Aspergillus terricola, Carica papaya and pseudomonodaceae). This paper reports an in vitro investigation, an in vivo study with rats and a clinical evaluation. It was found that crab collagenase has the highest proteolytic activity measured with respect to fibrin clot and necrotic eschar in vitro although its caseinolytic activity is the lowest. A clinical trial involving 21 patients with leg ulcers confirmed the clinical efficiency of crab collagenase compared with trypsin/ chymotrypsin. The results suggest that crab collagenase is useful in wound debridement.

Immunohistochemical study of purulent wounds treated with King crab collagenase.

Immunohistochemical study of tissues from purulent wounds in rats after treatment with the collagenase isolated from the King crab Paralithodes camtschatica was undertaken. The enzymotherapy resulted in a rapid and efficient removal of necrotic debris. It was accompanied by fibrin elimination from the wound bed and subsequent formation of new capillaries. Cellular fibronectin with ED-A sequence was identified in the newly formed granulation tissue, which points to its active synthesis in situ. Polyclonal antibodies against two isozymes of the crab collagenolytic protease were obtained. By their use it was shown that, after application of the collagenase, both isozymes accumulated in fibrin deposits at the wound bed but did not penetrate adherent granulation tissue.

[The immunohistochemical study of suppurative wounds in rats following the application of collagenase from the crab Paralithodes camtschatica]. / Immunogistokhimicheskoe izuchenie gnoinykh ran u krys posle applikatsii kollagenazy kraba Paralithodes camtschatica.

Immunohistochemical study of tissues of purulent wounds in rats after application of the collagenase isolated from the king crab Paralithodes camtschatica has been undertaken. The enzyme therapy resulted in a rapid and efficient removal of necrotic debris. It was accompanied by fibrin elimination from the wound bottom and subsequent formation of new capillaries. Cellular fibronectin with ED-A sequence was identified in the newly formed granulation tissue, which points to its active synthesis in situ. Detection of type I collagen in granulation tissue revealed that wound treatment with crab collagenase had no impact on the development process of the tissue. Polyclonal antibodies against two isozymes of crab collagenolytic protease were obtained. It was shown that after application of both isozymes of the collagenase were accumulated in fibrin deposits at the wound bottom but not penetrated in adherent granulation tissue. These processes underlie the therapeutic effect of the crab collagenase.

[A morphological evaluation of the action of collagenase from the king crab Paralithodes camtschatica on the experimental wound process]. / Morfologicheskaia otsenka vozdeistviia kollagenazy kamchatskogo kraba Paralithodes camtschatica na ranevoi protsess v éksperimente.

The paper presents the results of experimental morphological evaluation of the effect of a new proteolytic enzyme collagenase of crab Paralithodes camtschatica on wound healing in infected and aseptic rabbit wounds. The enzyme was applied on wounds using gauze and gelevin. The findings show that this protease is highly effective for debridement of infected wounds, the effect increasing at gelevin addition. To reach maximal therapeutical effect and diminish the inhibition effect of collagenase on granulation tissue, it is recommended to reduce the dose of protease during debridement process. Clinical application of crab collagenase must be individual as well as duration of wound enzymotherapy.