scholarly journals Overcoming the Inflammatory Stage of Non-Healing Wounds: In Vitro Mechanism of Action of Negatively Charged Microspheres (NCMs)

Nanomaterials ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 1108
Author(s):  
Edorta Santos-Vizcaino ◽  
Aiala Salvador ◽  
Claudia Vairo ◽  
Manoli Igartua ◽  
Rosa Maria Hernandez ◽  
...  

Negatively charged microspheres (NCMs) represent a new therapeutic approach for wound healing since recent clinical trials have shown NCM efficacy in the recovery of hard-to-heal wounds that tend to stay in the inflammatory phase, unlocking the healing process. The aim of this study was to elucidate the NCM mechanism of action. NCMs were extracted from a commercial microsphere formulation (PolyHeal® Micro) and cytotoxicity, attachment, proliferation and viability assays were performed in keratinocytes and dermal fibroblasts, while macrophages were used for the phagocytosis and polarization assays. We demonstrated that cells tend to attach to the microsphere surface, and that NCMs are biocompatible and promote cell proliferation at specific concentrations (50 and 10 NCM/cell) by a minimum of 3 fold compared to the control group. Furthermore, NCM internalization by macrophages seemed to drive these cells to a noninflammatory condition, as demonstrated by the over-expression of CD206 and the under-expression of CD64, M2 and M1 markers, respectively. NCMs are an effective approach for reverting the chronic inflammatory state of stagnant wounds (such as diabetic wounds) and thus for improving wound healing.

2015 ◽  
Vol 51 (2) ◽  
pp. 415-427 ◽  
Author(s):  
Juliana Neves Rodrigues Ract ◽  
Fabiana Andreia Schäfer De Martini Soares ◽  
Hosana Gomes Rodrigues ◽  
José Ricardo Bortolon ◽  
Gilson Masahiro Murata ◽  
...  

<p>Two oil blends (sunflower/canola oils 85/15 (BL1) and canola/linseed oils 70/30 (BL2)), were prepared and enzymatically interesterified to be applied to surgically-induced wounds in rats. Following surgery, the animals were submitted to the Treatment with Physiological Saline (TPS) (control group), Blends (TBL), and Structured Lipids (TSL). The control group (TPS) received physiological saline solution for 15 days. In TBL, BL1 was administered during the inflammation phase (days 0-3) and BL2 in the tissue formation and remodeling phase (days 4-15). In TSL, Structured Lipid 1 (SL1) and Structured Lipid 2 (SL2) were used instead of BL1 and BL2, respectively. The aim of this study was to compare wound closure evolution among rats treated with the blends or structured lipids versus control rats treated with physiological saline. The wound healing process was evaluated by measuring the wound areas along the treatments and the concentrations of cytokines. An increase in the areas of wounds treated with the blends and structured lipids in the inflammatory phase was observed, followed by a steeper closure curve compared to wounds treated with physiological saline. The changes observed during the inflammatory phase suggest a potential therapeutic application in cutaneous wound healing which should be further investigated.</p>


2018 ◽  
Vol 4 (2) ◽  
pp. 109
Author(s):  
Mufimah Mufimah ◽  
Uti Rusdian Hidayat ◽  
Ichsan Budiharto

Abstract: Efectiveness Gel Extract Of White On The Process Of Healing Inflamation Phase Heating. The inflammatory phase is a favorable body response as a protection mechanism. In the process of wound healing becomes a very important phase. Management of inflammation that is often used Non-Steroid Anti-Inflammatory class of salicylates on the skin that have side effects. The content of allicin in garlic can be used for problems that begin with the inflammatory phase. The use of gel from garlic extract is also easier to use and easier to clean. The study aim to determine the effectiveness of garlic extract gel to process wound inflammatory phase healing. This research is an experimental research with pre and post test with control group method with 24 samples. Conducted injury to the back area of rat length of wound 1 cm, depth to dermis. Conducted wound care, given gel extract of garlic concentration of 20%, 40%, 80% of the control using 0.9% NaCl compress. Using Kruskal Wallis test and Anova oneway showed concentration of 20%, 40%, and 80% of sig <0,05 ie 0.00. It was concluded that 20%, 40%, 80% garlic extract gel was effective against inflammatory wound healing process. The use of garlic extract gel is more effective in the wound inflammatory wound healing process.Abstrak: Efektivitas Gel Ekstrak Bawang Putih  terhadap Proses Penyembuhan Luka Fase Inflamasi.  Fase inflamasi merupakan respon tubuh yang menguntungkan sebagai mekanisme perlindungan. Pada proses penyembuhan luka menjadi fase yang sangat penting. Penatalaksanaan inflamasi yang sering digunakan Anti-Inflamasi Non Steroid golongan salisilat pada kulit yang memiliki efek samping. Kandungan zat allicin pada bawang putih dapat dimanfaatkan untuk masalah yang diawali dengan fase inflamasi. Pemanfaatan gel dari ekstrak bawang putih pun dalam penggunaannya lebih mudah diabsorsi dan mudah dibersihkan. Penelitian ini bertujuan untuk mengetahui efektifitas gel ekstrak bawang putih terhadap proses penyembuhan luka fase inflamasi. Penelitian ini merupakan penelitian eksperiment dengan metode pre and post test with control grup dengan jumlah sampel sebanyak 24 ekor tikus. Dilakukan perlukaan pada daerah punggung tikus panjang luka 1 cm, kedalaman sampai dermis. Dilakukan perawatan luka, diberi gel ekstrak bawang putih konsentrasi berbeda yaitu 20%, 40%, 80%  kontrol menggunakan kompres NaCl 0,9%. Hasil uji Kruskal Wallis dan Anova oneway menunjukkan konsentrasi 20%, 40%, dan 80%  nilai sig <0,05 yaitu 0,00. Disimpulkan bahwa 20%, 40%, 80% gel ekstrak bawang putih efektif terhadap proses penyembuhan luka inflamasi. Penggunaan gel ekstrak bawang putih lebih efektif dalam proses penyembuhan luka inflamasi luka.   Disimpulkan bahwa 20%, 40%, 80% gel ekstrak bawang putih efektif terhadap proses penyembuhan luka inflamasi. Penggunaan gel ekstrak bawang putih lebih efektif dalam proses penyembuhan luka inflamasi luka.


2020 ◽  
Vol 5 (1) ◽  
pp. 21
Author(s):  
Desty Ayu Dwiyanti ◽  
Irham Taufiqurrahman ◽  
Didit Aspriyanto

Background: Ethanol extract of binjai leaves contain bioactive compounds in the form of flavonoid and saponin which is potential as anti-inflammatory, so it will be able to control the wound healing process. Binjai leaves extract can affect the number of neutrophil on inflammatory phase, as it is characterized by neutrophil cells infiltration to the wound tissue, so its number will increase at the beginning of inflammatory phase and decrease on the 3rd day. Objective: To prove the effect of binjai leaves extract in 1 mg/mL, 2 mg/mL, and 3 mg/mL which given topically to the number of neutrophil in day 1, 3, and 5 of wound incision on the back of male mice. Methods: The study design was a pure laboratory experimental study with post-test only with control group design which involved 36 mice divided into 4 groups: negative control group design which was given BR2 feed, treatment group which was given binjai leaves extract in 1 mg/mL, 2 mg/mL, and 3 mg/mL. Results: The result of One-way ANOVA showed that there was significant difference of all groups in day 1 (p=0.000), day 3 (p=0.001), and day 5 (p=0.000). Post-Hoc LSD test showed that there was significant difference (p<0.05) between the negative control group and treatment groups. Conclusion: Binjai leaves extract has an effect to increase the number of neutrophil in day 1 and decrease the number of neutrophil in day 3 and 5 on the healing of back wound incision in mice.


2014 ◽  
Vol 21 (1) ◽  
pp. 13
Author(s):  
Endah Kusumastuti ◽  
Juni Handajani ◽  
Heni Susilowati

Inflamasi merupakan respon alami tubuh terhadap adanya kerusakan jaringan. Salah satu medikamen untuk mengatasi inflamasi adalah antiinflamasi non steroid (AINS). Penggunaan AINS mempunyai beberapa efek samping dan dalam beberapa hal penggunaan tanaman obat dinilai lebih aman. Rosela merupakan salah satu tanaman obat yang mempunyai potensi sebagai antiinflamasi. Penelitian ini bertujuan untuk mengetahui efek pemberian sistemik ekstrak etanolik rosela terhadap ekspresi COX-2 dan jumlah neutrofil fase inflamasi pada proses penyembuhan luka. Bunga rosela didapatkan dari perkebunan di Dusun Bulusari Desa Pojok Kecamatan Tarokan Kabupaten Kediri Jawa Timur. Pembuatan ekstrak rosela dilakukan di LPPT unit I UGM Yogjakarta dengan cara perkolasi. Tikus putih galur Wistar sebanyak 36 ekor diberi perlukaan dengan punch biopsi ɵ 3 mm pada mukosa bukal. Subjek dibagi menjadi 3 kelompok, masing-masing kelompok 12 ekor tikus. Pembagian kelompok terdiri dari kontrol negatif (saline), kontrol positif (ibuprofen 20 mg/kg BB) dan perlakuan (ekstrak rosela 500 mg/kg BB). Pemberian minum sesuai kelompoknya sehari sekali selama 4 hari. Pada hari ke-1, ke-2, ke-3 dan ke-4 tikus dikorbankan lalu jaringan mukosa yang mengalami perlukaan dibuat preparat histologis. Pewarnaan Hematoksilin Eosin (HE) dilakukan untuk mengamati jumlah neutrofil. Ekspresi COX-2 diamati pada preparat dengan pewarnaan imunohistokimia menggunakan rabbit polyclonal antibody COX-2 (Lab Vision, USA). Jumlah neutrofil dan ekspresi COX-2 dihitung di bawah mikroskop cahaya lalu data dianalisi menggunakan ANAVA dan LSD. Hasil penelitian menunjukkan bahwa ekspresi COX-2 dan jumlah neutrofil lebih rendah pada kelompok perlakuan dibanding kontrol. Pengamatan klinis pada hari ke-4 juga tampak luka seluruh subjek telah menutup sempurna setelah pemberian minum rosela. Disimpulkan bahwa ekstrak etanolik rosela mempunyai kemampuan menghambat ekspresi COX-2 dan menurunkan jumlah neutrofil sehingga dapat digunakan sebagai bahan anti-inflamasi. Expression of COX-2 and The Number of Neutrophil in Inflammation stage of Wound Healing Process after Systemic Administration of Ethanolic Extract Rosela. Inflammation is an initial stage of body’s natural response to tissue damage.The use  empirically plants often used for traditional medicine because it is easily found in the community and fewer side effects. Flavanoid presence of roselle (Hibiscus sabdariffa) is thought to have anti inflammatory effects. This study aimed to know the effect of systemic administration of Roselle ethanolic extract toward COX-2 expression and neutrophils number in the inflammatory phase of wound healing processes. Roselle was obtained from plantations in Bulusari hamlet, Tarokan, Kediri, EastJava. Making roselle extract was performed in LPPT unit 1 UGM Yogyakarta by percolation ways. Wistar rats were given a total of 36 injuries with ɵ 3 mm punch biopsy of the buccal mucosa. Subjects were divided into three groups, each group of 12 rats. The division consists of the negative control group (saline), positive control (ibuprofen 20 mg/kg) and treatment (roselle extract 500 mg/kg). Giving drink suitable group once daily for four days. On day 1, the 2nd, 3rd and fourth rats were sacrificed, and mucosal tissue injury was made histological preparat. Hematoxylin eosin staining (HE) was performed to observe the number of neutrophils. COX-2 expression was found in preparations for immunohistochemical staining using rabbit polyclonal COX-2 antibody (Lab Vision, USA). The number of neutrophils and expression of COX-2 is calculated under a light microscope data were analyzed using Two-way ANOVA and LSD. The results showed that the expression of COX-2 and neutrophil number were least in the treatment group compared to the control. Clinical observation on day four also appears around the wound has completely closed the subject after administration of roselle drink. It was concluded that the ethanolic extract of roselle can inhibit COX-2 expression and decrease the number of neutrophils that can be used as an anti-inflammatory ingredient. 


2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Zhou Xu ◽  
Bin Deng ◽  
Xuewen Wang ◽  
Jie Yu ◽  
Zhuobin Xu ◽  
...  

Abstract Background Diabetic wound healing remains a challenge because of its susceptibility to drug-resistant bacterial infection and its persistent proinflammatory state. Switching from proinflammatory M1 macrophages (Mφs) to proregenerative M2 dominant Mφs in a timely manner accelerates wound healing by coordinating inflammatory, proliferative, and angiogenic processes. Methods We propose a sequential photothermal antibacterial and subsequent M2 Mφ polarization strategy based on nanofibers (NFs) consisting of polydopamine (PDA) coating on curcumin (Cur) nanocrystals to treat Methicillin-resistant Staphylococcus aureus (MRSA)-infected diabetic wounds. Results The PDA/Cur NFs showed excellent photothermal conversion and antibacterial effects due to the PDA shell under laser irradiation, consequently resulting in the release of the inner Cur with the ability to promote cell proliferation and reinforce the M2 Mφ phenotype in vitro. In vivo studies on MRSA-infected diabetic wounds showed that PDA/Cur NFs not only inhibited MRSA infection but also accelerated the wound regeneration process. Furthermore, the NFs displayed the ability to promote the M2 Mφ phenotype with enhanced collagen deposition, angiogenesis, and cell proliferation. Conclusion Overall, the NFs displayed great potential as promising therapeutics for healing infected diabetic wounds through a sequential photothermal antibacterial and M2 Mφ polarization strategy. Graphical abstract


2017 ◽  
Vol 9 (3) ◽  
pp. 306-311 ◽  
Author(s):  
DESAK MADE MALINI ◽  
MADIHAH MADIHAH ◽  
FITRI KAMILAWATI ◽  
NINING RATNINGSIH ◽  
KARTIAWATI ALIPIN ◽  
...  

Malini DM, Madihah, Kamilawati F, Ratningsih N, Alipin K, Iskandar J. 2017. Topical treatment of ointment containing ethanol extract of Archidendron pauciflorum fruit peel on the wound healing in streptozotocin-induced diabetic mice. Nusantara Bioscience 9: 306-311. Diabetic wounds lead to severe tissue damage and are difficult to cure. One alternative medicine known well by local Indonesian communities to treat diabetic wounds is the fruit peel of djengkol. This study aimed to evaluate the ointment containing ethanol extract of djengkol fruit peel to accelerate wound healing process in the skin of streptozotocin-induced diabetic mice. The method was experimental using completely randomized design with six treatments and four replications. Diabetes was induced by intraperitoneal injection of streptozotocin 180 mg/kg BW. Mice with blood glucose level ≥150 mg/dL were used for diabetic mice models. The incision wound created at dorsolateral region of shaven skin at ±1 cm2 using sterile scissors. The ointment containing extract was applied topically to the diabetic mice wounds at concentration of 5%, 10% and 15%, as well as Betadine® as the reference group. The ointment basis was applied to the wound of diabetic mice as a positive control and to the wound of non-diabetic mice as a negative control. The treatment was done twice a day for 14 days. The results showed that topical application of ointment containing ethanol extract of djengkol fruit peel at concentration of 10% gave significant recovery (p<0.05) on the wounded skin by the enhancement of re-epithelization and granulation tissue, as well as the increase of capillary number and collagen density which were higher than other treatments and comparable to negative control group. It was concluded that the topical application of ointment containing ethanol extract of djengkol fruit peel can support the healing of diabetic wounds.


2014 ◽  
Vol 13 (3) ◽  
pp. 176
Author(s):  
Arisa Izzaty ◽  
Nurdiana Dewi ◽  
Dewi Indah Noviana Pratiwi

Haruan is river-water fish usually consumed by South Kalimantan people. Haruan extract contains albumin, Zn, Fe,Cu and unsaturated fatty acid which can act as antiinflammatory substances and accelarate wound healing. The aim ofthis study is to examine effectiveness of haruan extract on lymphocyte count in infammatory phase of wound healingprocess and compare it to ibuprofen. This study used true experimental method with post-test only control group designand simple random sampling. Samples were divided into 5 treatment groups:which were given the extract for 7 sevendays: 25% haruan extract group, 50% haruan extract group, 100% haruan extract group, negative control K(-) usingaquadest, positive control K(+) given ibuprofen suspension with dosage of 20mg/kg weight/day. Lymphocyte cells wereobserved using microscope and counted per field of view. Lymphocyte count reached its peak on day 5. Lymphocytecount means in each group were 4,2 ± 0,566; 1± 0,000; 3,8 ± 2,546; 7,1 ± 0,141; dan 7,5 ± 3,818 respectively. Two way ANOVA and post hoc LSD tests showed that lymphocyte count significantly decreased in 50% and 100% treatmentgroups (p<0,05) and has equal effect with ibuprofen in decreasing lymphocyte count. This study concluded that haruanextract significantly decreased lymphocyte count in wound healing process.


2020 ◽  
Vol 133 (19) ◽  
pp. jcs235838
Author(s):  
Peilang Yang ◽  
Xiqiao Wang ◽  
Di Wang ◽  
Yan Shi ◽  
Meng Zhang ◽  
...  

ABSTRACTBesides regulating glucose levels, insulin has been reported to participate actively in many other functions, including modulating inflammatory reactions. In this study we investigated how topical insulin application would affect the diabetic wound healing process. We found that the excessive expression of insulin-degrading enzyme led to insufficient insulin levels in diabetic skin during wound healing, which ultimately reduced the recovery rate of diabetic wounds. We confirmed that topical insulin application could reverse the impaired inflammation reaction in the diabetic wound environment and promote healing of diabetic wounds. Our study revealed that insulin promoted apoptosis of neutrophils and subsequently triggered polarization of macrophages. Both in vivo and in vitro studies verified that insulin re-established phagocytosis function and promoted the process of phagocytosis-induced apoptosis in neutrophils. Furthermore, we found that insulin treatment also promoted efferocytosis of the apoptosed neutrophils by macrophages, and thus induced macrophages to change their polarization state from M1 to M2. In conclusion, our studies proved that the exogenous application of insulin could improve diabetic wound healing via the restoration of the inflammatory response.


Cells ◽  
2019 ◽  
Vol 8 (8) ◽  
pp. 917
Author(s):  
Rafael Ballesteros-Cillero ◽  
Evan Davison-Kotler ◽  
Nupur Kohli ◽  
William S. Marshall ◽  
Elena García-Gareta

Due to great clinical need, research where different biomaterials are tested as 3D scaffolds for skin tissue engineering has increased. In vitro studies use a cell suspension that is simply pipetted onto the material and cultured until the cells migrate and proliferate within the 3D scaffold, which does not mimic the in vivo reality. Our aim was to engineer a novel biomimetic in vitro model that mimics the natural cell infiltration process occurring in wound healing, thus offering a realistic approach when pre-screening and testing new skin substitutes. Our model consists of porous membrane cell culture inserts coated with gelatin and seeded with human dermal fibroblasts, inside which two different commercially available dermal substitutes were placed. Several features relevant to the wound healing process (matrix contraction, cell infiltration and proliferation, integration of the biomaterial with the surrounding tissue, and secretion of exogenous cytokines and growth factors) were evaluated. Our results showed that cells spontaneously infiltrate the materials and that our engineered model is able to induce and detect subtle differences between different biomaterials. The model allows for room for improvements or “adds-on” and miniaturization and can contribute to the development of functional and efficient skin substitutes for burns and chronic wounds.


2012 ◽  
pp. 241-250 ◽  
Author(s):  
P. GÁL ◽  
T. VASILENKO ◽  
I. KOVÁČ ◽  
M. KOSTELNÍKOVÁ ◽  
J. JAKUBČO ◽  
...  

Previously, we found that treatment of cutaneous wounds with Atropa belladonna L. (AB) revealed shortened process of acute inflammation as well as increased tensile strength and collagen deposition in healing skin wounds (Gál et al. 2009). To better understand AB effect on skin wound healing male Sprague-Dawley rats were submitted to one round full thickness skin wound on the back. In two experimental groups two different concentrations of AB extract were daily applied whereas the control group remained untreated. For histological evaluation samples were removed on day 21 after surgery and stained for wide spectrum cytokeratin, collagen III, fibronectin, galectin-1, and vimentin. In addition, in the in vitro study different concentration of AB extract were used to evaluate differences in HaCaT keratinocytes proliferation and differentiation by detection of Ki67 and keratin-19 expressions. Furthermore, to assess ECM formation of human dermal fibroblasts on the in vitro level fibronectin and galectin-1 were visualized. Our study showed that AB induces fibronectin and galectin-1 rich ECM formation in vitro and in vivo. In addition, the proliferation of keratinocytes was also increased. In conclusion, AB is an effective modulator of skin wound healing. Nevertheless, further research is needed to find optimal therapeutic concentration and exact underlying mechanism of action.


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