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Riordan et al. J Transl Med (2015) 13:242
DOI 10.1186/s12967-015-0608-8

Case Report:

Case report of non‑healing surgical
wound treated with dehydrated human
amniotic membrane

Neil H Riordan1,2*, Ben A George2, Troy B Chandler2 and Randall W McKenna2


ABSTRACT

INTRODUCTION: Non-healing wounds can pose a medical challenge as in the case of vasculopathic venostasis resulting in a surgical ulcer. When traditional approaches to wound care fail, an amniotic patch containing mesenchymal stem cells may present a viable solution for therapeutic and efficient wound care.

BACKGROUND: Morselized amniotic AlphaPATCH patches contain concentrated molecules of PGE2, WNT4, and GDF-11, which have angiogenic, trophic, and anti-inflammatory effects on tissues that may be useful in the efficient healing of wounds.

AIM – CASE REPORT: We present a case of a severe non-healing surgical wound with sympathetic effusion in a 78- year-old male who underwent a right total knee arthroplasty procedure.

MATERIALS AND METHODS: In the OR, the joint effusion was drained, and 65cc of bloody synovial fluid was removed from the patient’s right knee. STAT gram stain of the aspirate revealed negative results. The wound was then debrided and irrigated with three liters of double antibiotic solution under pulse lavage. Two dry amniotic AlphaPATCH patches (4cm x 4cm) were placed over the wound with Acticoat applied on top.

RESULTS: At the two-week follow-up visit (following the incision and drainage of the wound dehiscence and application of the amniotic AlphaPATCH patches), a central scab formed in the middle of the wound dehiscence area (figure B). At the four-week follow-up visit, the wound dehiscence area completely scabbed over with no open areas left (figure C). At the eight-week follow-up visit, the scab had just fallen off, and the wound was healing well with immature skin representing the size of a penny (figure D). At the ten-week follow-up visit, the wound was completely healed (figure E).

DISCUSSION/CONCLUSION: Morselized amniotic AlphaPATCH patches containing mesenchymal stem cells can successfully treat non-healing wounds and offer a viable and more effective alternative to current traditional means of wound care management.


BACKGROUND

Wound healing is a complex process whereby multiple cell types, growth factors, and extra-cellular proteins interact to repair a disruption in the dermal and epidermal layers of the skin. In some cases, the mechanism behind said repair fails to restore the integrity of the injured tissue in a timely manner, delaying the progression of inflammatory, proliferative, and remodeling phases of healing[1]. The resulting chronic, non-healing wound is vulnerable to infection, may cause pain, reduce quality of life, and become a burden on the healthcare system[2]. Non-healing wounds, frequently seen in older patients[3], are associated with certain conditions such as diabetes, obesity, and rheumatoid arthritis, but may also occur following acute trauma or surgical intervention[4, 5]. It is estimated that 1-2% of patients in developed countries will experience non-healing wounds in their lifetime[6], and certain types of chronic wounds are estimated to account for billions of dollars of treatment costs in the United States[3].

An adequate progression of wound healing[1, 7, 8] begins with the secretion of growth factors such as Transforming Growth Factor beta (TGF-β)[9], as well as Fibroblast (FGF), Endothelial (EGF), Platelet Derived (PDGF) and Vascular Endothelial (VEGF) Growth Factors[10]. Neutrophils attracted by PDGF signals clear excess bacteria at the site with the aid of monocytes – later transformed to macrophages. Macrophages regulate the production of TGF-β, which in turn stimulates migration and proliferation of fibroblasts as well as epithelialization[11]. Extracellular matrix and granulation tissue begin forming as fibroblasts secrete fibronectin and collagen precursors concurrently with VEGF-stimulated angiogenesis which carries oxygen and nutrients to the injured site[12]. Finally, the collagen structure in the wound area matures and reassembles into a tighter structure with greater tensile strength. An interruption or delay at any stage of this complex process results in a non-healing wound[2, 13, 14].

The first line of treatment for non-healing wounds is to keep the injured area clean and to debride any infected or necrotic tissue; antibiotics may be prescribed to address bacterial infection. Certain types of chronic wounds can be treated with compression therapy or with negative pressure wound devices[15], whereby a vacuum drains wound fluid and increases blood flood to the area. In the case of more unmanageable non-healing wounds, a skin graft may be considered[16, 17]: an autologous transplant is less likely to be rejected but is limited by the size of the wound and the extent of available healthy tissue to replace the damaged one. Conversely, an allograft from cadaveric skin may cover a larger area but can trigger an immune reaction against the donor tissue. Tissue bioengineering represents another viable alternative for non-healing wounds[18-20], whereby in vitro-grown cells are placed on an artificial scaffold (often collagen-based) at the wound site to encourage cell migration to the site and tissue repair, eventually restoring functionality to the area. However, this high-cost technique cannot fully replicate the biological and cellular characteristics of true skin[18], notably with regards to angiogenesis at the wound site, and has not been shown to be significantly more efficient than other treatments. Standard treatment for wound care still fails in an estimated 50% of cases[13].

Cell therapy has emerged as a non-invasive, viable alternative for treating non-healing wounds, particularly using mesenchymal stem cells (MSCs)[21]. MSCs are pluripotent cells capable of differentiation into various cell types[22], and may be isolated from bone marrow, adipose tissue, umbilical cord, placenta and embryonic tissue[23]. MSCs have a trophic effect that stimulates tissue
repair[24] and angiogenesis, as well as the ability to move towards damaged tissue and to regulate inflammation at the site[25]. The immunomodulatory properties of MSCs (MHC I+, MHC II-, CD40-, CD80-, CD86- and CD44+, Cd73+, CD90+, CD105+)[25] prevent immunosuppression and allow recruitment of more macrophages, thereby stimulating the release of TGF-β, EGF, FGF, PDGF, and VEGF among others[14, 24, 26]. The angiogenic, anti-inflammatory, and trophic effects of MSCs, therefore, hasten the progression of healing and remodeling and yield many applications in clinical settings.

MSCs have been used to treat non-healing wounds[27, 28], notably Bone Marrow-derived MSCs[29], with reports of full healing at 21 days[30], complete healing within 8 weeks[31], no wound recurrence[30], and other improvements[32, 33], Similarly, adipose-derived MSCs have yielded satisfactory outcomes for non-healing wounds[34, 35]. Amniotic products and umbilical chord tissue rich in MSCs may also be used for wound and burn treatments. These products are more accessible[36], larger in numbers, and hold more proliferative capacity[37, 38] relative to adult MSCs. Furthermore, since this neonatal tissue is disposed of after birth, it does not present the ethical problems associated with the use of embryonic material[36]. Of particular interest is amnion, the inner-membrane holding the embryo sac[39, 40]. This tissue may be cryo-preserved after birth to retain the qualities of amniotic membrane and the growth factors secreted in the amniotic environment such as prostaglandin and WNT4 (which reduce inflammation and promote healing). Amniotic membrane dressing[41] has been shown to perform significantly better than conventional treatment. One study reports 81% epithelialization in 2 weeks, no infection in 80% of cases, drop in exudation, healthy granulation tissue, and decrease of pain[42]. In another study, wound closure was greater than 40% with a reduction in mean size in the group treated with amniotic membrane compared to the control group treated with compression therapy[43].

This case report presents a patient with a non-healing wound following a knee replacement surgery. This rare complication of arthroplasty[44, 45] is estimated to occur in 0.33% of cases in the 30 days following an operation, and 5.3% of cases need subsequent surgeries in the following 2 years[46].


CASE REPORT

A 78-year-old male with history of prostate cancer, COPD, and smoking a pack of cigarettes daily (for four decades) presented for a follow-up visit in the clinic 17 days status post right total knee arthroplasty. During this visit, dermal staples were removed from the surgical incision. The mid portion of the patient’s incision showed some wound dehiscence, representing the same location in which the patient reported a previous sebaceous cyst “that had to heal from the inside out.” As antibiotic prophylaxis, Cleocin 300 mg was prescribed to be taken four times daily while Talwin was prescribed one tablet every four hours as needed for pain control. The dehisced area was thoroughly cleaned with alcohol and triple antibiotic while the remainder of the wound was Steri-Stripped.

One week later, the patient presented to the clinic for follow-up of the wound dehiscence. Upon physical examination, the wound appeared macerated and white with initial signs of necrosis. The wound was cleaned with alcohol and compressed with Ace bandage. The patient has continued to take Cleocin antibiotic regimen.

Eleven days later, the patient returned to the clinic to recheck the wound. The wound showed slow healing with no significant drainage. To expedite the healing process, three Monocryl sutures were used to close the wound.

One week later (or 1-1/2 months status post total knee arthroplasty), the patient returned to the clinic reporting that the three sutures popped out while he flexed his knee during physical therapy. The non-healing wound displayed a sympathetic effusion consistent with vasculopathic venostasis resulting in a surgical ulcer. The wound, however, was not erythematous, hot, or tender to palpation. The patient was scheduled the following day in the OR for drainage of joint effusion with gram stain, knee wound irrigation with pulse lavage, and application of amniotic dry patch to wound.


PROCEDURE

The right lower extremity was sterilely prepped and draped in the usual fashion, roughly two months status post total knee arthroplasty. The joint effusion was drained, and 65cc of bloody synovial fluid was removed from the patient’s right knee. STAT gram stain of the aspirate revealed negative results. The wound was then debrided and irrigated with three liters of double antibiotic solution under pulse lavage. Two dry amniotic AlphaPATCH patches (4cm x 4cm) were placed over the wound with Acticoat applied on top. Then 4×4’s, Webril, and Ace wrap were applied to the knee. The patient tolerated the procedure well and was transferred to the recovery room in stable condition.


RESULTS

At the two-week follow-up visit (following the incision and drainage of the wound dehiscence and application of the amniotic AlphaPATCH patches), a central scab has formed in the middle of the wound dehiscence area (figure B). At the four-week follow-up visit, the wound dehiscence area has completely scabbed over with no open areas left (figure C). At the eight-week follow-up visit, the scab has just fallen off, and the wound is healing well with immature skin representing the size of a penny (figure D). At the ten-week follow-up visit, the wound has completely healed (figure E), patient demonstrated full knee ROM (120 degrees of flexion and 180 degrees of extension), and patient has been released from orthopaedic care.

CaseReportHealing

DISCUSSION

Although the 78-year-old patient demonstrated excellent results in ROM and overall decreased pain levels following right total knee arthroplasty, the non-healing status of the surgical wound at even six weeks status post surgery posed great concern. The vasculopathic venostasis resulting in a surgical ulcer left very little option than to perform dehiscence wound irrigation with pulse lavage and to apply amniotic AlphaPATCH dry patches to the wound following confirmation of gram negative status of the aspirate.

The amniotic AlphaPATCH patch, a morselized amniotic tissue allograft, contains the molecules PGE2, WNT4, and GDF-11. Mesenchymal stem cells (MSCs) secrete PGE2, or Prostaglandin E2, in response to injury. This molecule inhibits fibrosis by way of limiting fibroblast proliferation, migration, collagen secretion, and transforming growth factor (TGF)-induced myofibroblast that can spur fibroblast proliferation[47]. PGE2 also enhances the wound healing process and angiogenesis[48]. WNT4, a protein, drives wound healing by way of wound re-epithelialization and cell proliferation[49]. The creation of new tissue materializes by multiple methods including new blood vessel formation, a critical element for normal wound healing[50]. GDF-11, or growth/differentiation factor 11, has been identified as one of the key molecules propelling the regeneration of skeletal muscle, cardiac muscle, and nervous tissue in aged mice as a result of heterochronic parabiosis, a procedure used to connect the circulatory system of a young mouse to that of an older mouse[51].

The two (MSC-based) amniotic AlphaPATCH patches applied on the patient’s wound dramatically accelerated the wound healing process. The dehisced surgical wound that showed no sign of healing after 1-1/2 months post total knee replacement surgery, demonstrated a central scab formation in the middle of the wound dehiscence area (figure B) only after two weeks of AlphaPATCH patch application. After eight more weeks, the wound was completely healed and the patient was released from orthopaedic care to assume high levels of physical activity and activities of daily living. Although more studies are warranted to further substantiate the therapeutic benefits of this treatment, we suggest unreservedly that amniotic AlphaPATCH patch embodies a viable and more effective alternative to current traditional means of wound care management.


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