次氯酸对骨科内固定物表面细菌生物膜的清除作用

梁金正 1 ，邵欣欣 2 ，杜少华 1 ，戴双武 1

文题释义：

次氯酸：化学式HClO，是氯气溶解于水时形成的一种一价氯的含氧弱酸。在工业上，次氯酸作为家用漂白剂和清洁剂中的活性成分而广为人知。在生物学系统中，次氯酸可被视为白细胞例如中性粒细胞和单核细胞产生的毒性最强、丰度最高的氧化剂。

细菌生物膜：也称为生物被膜，是指附着于有生命或无生命物体表面被细菌胞外大分子包裹的有组织的细菌群体，存在各种主要的生物大分子如蛋白质、多糖、DNA、RNA、肽聚糖、脂和磷脂等物质。细菌生物膜多细胞结构的形成是一个动态过程，包括细菌起始黏附、细菌生物膜发展和成熟扩散等阶段。细菌生物膜对抗生素和宿主免疫防御机制的抗性很强。

摘要

背景：骨科内固定物感染中形成的细菌生物膜是临床非常棘手的问题。次氯酸作为一种高效消毒剂，可引起细菌生物膜破坏，可能对骨科内固定物感染治疗有帮助。

目的：研究次氯酸溶液体外浸泡不同时间及灌注对骨科内固定物表面细菌生物膜的清除作用。

方法：将金黄色葡萄球菌和铜绿假单胞菌分别在骨科钢板内固定物上定植3 d后形成细菌生物膜，扫描电镜下确认细菌生物膜已形成。采用次氯酸溶液分别浸泡已形成两种细菌生物膜的内固定物0，5，10，20 min及20 min+灌注6 L次氯酸和生理盐水的混合溶液(每3 L生理盐水加入120 mL次氯酸)，扫描电镜下观察细菌生物膜形态，评估次氯酸溶液对内固定物细菌生物膜的清除效果。

结果与结论：①铜绿假单胞菌：扫描电镜下可见，未经次氯酸溶液浸泡的内固定物上的细菌生物膜斑块面积大且较厚，菌落密集集中；经过次氯酸浸泡5，10，20 min后，细菌生物膜斑块面积逐渐减小并变薄，次氯酸溶液浸泡20 min+灌注6 L次氯酸和生理盐水的混合溶液可清除细菌生物膜；②金黄色葡萄球菌：扫描电镜下的表现与铜绿假单胞菌生物膜一致；③结果表明：延长浸泡时间或联合灌注可增强次氯酸对内固定物细菌生物膜的清除作用；次氯酸溶液浸泡足够时长可降解破坏内固定物上定植的细菌生物膜，清除菌落，从而达到消毒目的。

关键词：次氯酸；细菌生物膜；骨科手术；骨科内固定物；感染；消毒剂

¹南方医科大学第三附属医院骨科医学中心骨肿瘤科，广东省广州市 510630； ²中山大学附属第一医院麻醉科，广东省广州市 510080

第一作者：梁金正，男，1994 年生，广东省云浮市人，汉族，骨科住培医师，主要从事骨感染等方面研究。

通讯作者：戴双武，在读博士，主治医师，南方医科大学第三附属医院骨科医学中心骨肿瘤科，广东省广州市 510630 https://orcid.org/0000-0001-6824-5950 ( 梁金正 )

基金资助：广东省科技计划项目 (2020A1414050028)，项目负责人：戴双武

引用本文：梁金正，邵欣欣，杜少华，戴双武 . 次氯酸对骨科内固定物表面细菌生物膜的清除作用 [J]. 中国组织工程研究， 2022，26(15):2367-2371.

Liang Jinzheng1 , Shao Xinxin2 , Du Shaohua1 , Dai Shuangwu1

¹Department of Musculoskeletal Oncology, Center for Orthopedic Surgery, the Third Affiliated Hospital of Southern Medical University, Guangzhou 510630, Guangdong Province, China; ² Department of Anesthesiology, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China Liang Jinzheng, Resident of the Center for Orthopedic Surgery, the Third Affiliated Hospital of Southern Medical University, Guangzhou 510630, Guangdong Province, China

Corresponding author: Dai Shuangwu, MD candidate, Attending physician, Department of Musculoskeletal Oncology, Center for Orthopedic Surgery, the Third Affiliated Hospital of Southern Medical University, Guangzhou 510630, Guangdong Province, China

Abstract

BACKGROUND: Biofilm formation during the infection of orthopedic fixation devices is a very difficult clinical problem. Hypochlorous acid, as a highly effective disinfectant, can destroy the bacterial biofilm, which may be helpful for treating infection of orthopedic fixation devices.

OBJECTIVE: To study the scavenging effect of hypochlorous acid on bacterial biofilm on the surface of orthopedic fixation devices after soaking and perfusion for different time in vitro.

METHODS: Staphylococcus aureus and Pseudomonas aeruginosa were colonized on orthopedic steel fixation devices for 3 days, and then the bacterial biofilm formed, which was confirmed by scanning electron microscope. Orthopedic fixation devices with two kinds of bacterial biofilms were soaked in hypochlorous acid for 0, 5, 10, and 20 minutes, or perfused with 6 L of a mixed solution of hypochlorous acid and saline (every 3 L of saline with addition of 120 mL of hypochlorous acid) for 20 minutes. The morphology of bacterial biofilms was observed under scanning electron microscope, and the scavenging effect on the bacterial biofilm on the surface of orthopedic internal fixation devices was then evaluated.

RESULTS AND CONCLUSION: Pseudomonas aeruginosa: Under the scanning electron microscope, the bacterial biofilm plaques on the surface of the internal fixation device that was not soaked in hypochlorous acid solution were large and thick, and the colonies were densely concentrated. After 5, 10, and 20 minutes soaking in hypochlorous acid, the bacterial biofilm plaque area was gradually decreased and became thinner. Perfusion with 6 L of a mixture of hypochlorous acid and saline for 20 minutes could remove the bacterial biofilm. Staphylococcus aureus: The performance under the scanning electron microscope was  consistent with the biofilm of Pseudomonas aeruginosa. To conclude, prolonging the soaking time or a mixed perfusion can enhance the scavenging effect of hypochlorous acid on the bacterial biofilm on the surface of internal fixation devices. By soaking in hypochlorous acid for a sufficient time, the bacterial biofilm colonized on the surface of orthopedic internal fixation devices can be degraded and destroyed, thus achieving the purpose of disinfection.

Key words: hypochlorous acid; bacterial biofilm; orthopedic surgery; orthopedic internal fixation device; infection; disinfectant

Effects of Hypochlorous Acid and Hydrogen Peroxide Treatment on Bacterial Disinfection Treatments in Implantoplasty Procedures

Esteban Padulles-Gaspar ¹ , Esteban Padulles-Roig ² , Guillermo Cabanes ² , Román A. Pérez ³ , Javier Gil ^3,* and Begoña M. Bosch ^3,*

¹Facultad de Odontología, Universitat Internacional de Catalunya, C/Josep Trueta s/n, 08195 Barcelona, Spain

²Department of Implantology, University of La Salle, Madrid, EDE, C7Gaminedes 11, 28023 Madrid, Spain

³Bioengineering Institute of Technology, Universitat Internacional de Catalunya, 08195 Barcelona, Spain

^*Correspondence: 该Email地址已收到反垃圾邮件插件保护。要显示它您需要在浏览器中启用JavaScript。 (J.G.); 该Email地址已收到反垃圾邮件插件保护。要显示它您需要在浏览器中启用JavaScript。 (B.M.B.)

Abstract: One of the main problems in oral implantology today is peri-implantitis, which affects almost 20% of dental implants placed in patients. One of the most commonly used techniques to eliminate bacterial biofilm is the implantoplasty, that consists of the mechanical modification of the implant surface topography followed by treatments with chemical reagents for decontamination. In this study, the main aim is to evaluate the use of two different chemical treatments based on hypochlorous acid (HClO) and hydrogen peroxide (H2O2 ). For this purpose, 75 titanium grade 3 discs were treated with implantoplasty according to established protocols. Twenty-five discs were used as controls, 25 were treated with concentrated HClO and 25 were treated with concentrated HClO followed by treatment with 6% H2O2 . The roughness of the discs was determined using the interferometric process. Cytotoxicity with SaOs-2 osteoblastic cells was quantified at 24 and 72 h, whereas bacteria proliferation using S. gordonii and S. oralis bacteria was quantified at 5 s and 1 min of treatment. The results showed an increase in the roughness values, the control discs had an Ra of 0.33 µm and those treated with HClO and H2O2 reached 0.68 µm. Cytotoxicity was present at 72 h, together with a significant proliferation of bacteria. These biological and microbiological results can be attributed to the roughness produced by the chemical agents that triggered bacterial adsorption while inhibiting osteoblast adhesion. The results indicate that even if this treatment can decontaminate the titanium surface after implantation, the produced topography will generate an environment that will not favor long-term performance.

Keywords: hypochlorous acid; hydrogen peroxide; implantoplasty; periimplantitis

Effectiveness of Hypochlorous Acid to Reduce the Biofilms on Titanium Alloy Surfaces in Vitro

Chun-Ju Chen ¹ , Chun-Cheng Chen ^2,3,* and Shinn-Jyh Ding ^1,2,3,*

¹Institute of Oral Science, Chung Shan Medical University, Taichung City 402, Taiwan; 该Email地址已收到反垃圾邮件插件保护。要显示它您需要在浏览器中启用JavaScript。

²School of Dentistry, Chung Shan Medical University, Taichung City 402, Taiwan

³Department of Dentistry, Chung Shan Medical University Hospital, Taichung City 402, Taiwan

^*Correspondence: 该Email地址已收到反垃圾邮件插件保护。要显示它您需要在浏览器中启用JavaScript。 (C.-C.C.); 该Email地址已收到反垃圾邮件插件保护。要显示它您需要在浏览器中启用JavaScript。 (S.-J.D.);

Tel.: +886-6-2471-8668 (ext. 55255) (C.-C.C.); +886-6-2471-8668 (ext. 55529) (S.-J.D.)

Academic Editor: Ihtesham ur Rehman

Received: 13 April 2016; Accepted: 15 July 2016; Published: 19 July 2016

Abstract: Chemotherapeutic agents have been used as an adjunct to mechanical debridement for peri-implantitis treatment. The present in vitro study evaluated and compared the effectiveness of hypochlorous acid (HOCl), sodium hypochlorite (NaOCl), and chlorhexidine (CHX) at eliminating Gram-negative (E. coli and P. gingivalis) and Gram-positive (E. faecalis and S. sanguinis) bacteria. The effect of irrigating volume and exposure time on the antimicrobial efficacy of HOCl was evaluated, and a durability analysis was completed. Live/dead staining, morphology observation, alamarBlue assay, and lipopolysaccharide (LPS) detection were examined on grit-blasted and biofilm-contaminated titanium alloy discs after treatment with the three chemotherapeutic agents. The results indicated that HOCl exhibited better antibacterial efficacy with increasing irrigating volumes. HOCl achieved greater antibacterial efficacy as treatment time was increased. A decrease in antimicrobial effectiveness was observed when HOCl was unsealed and left in contact with the air. All the irrigants showed antibacterial activity and killed the majority of bacteria on the titanium alloy surfaces of biofilm-contaminated implants. Moreover, HOCl significantly lowered the LPS concentration of P. gingivalis when compared with NaOCl and CHX. Thus, a HOCl antiseptic may be effective for cleaning biofilm-contaminated implant surfaces.

Keywords: dental implant; hypochlorite acid; antiseptics; antimicrobial activity

Topical hypochlorite ameliorates NF-κB–mediated skin diseases in mice

Thomas H. Leung,^1,2 Lillian F. Zhang,¹ Jing Wang,¹ Shoucheng Ning,³ Susan J. Knox,³ and Seung K. Kim^1,4,5

¹Department of Developmental Biology, 2Department of Dermatology, 3Department of Radiation Oncology, 4Howard Hughes Medical Institute, and 5Department of Medicine, Oncology Division, Stanford University School of Medicine, Stanford, California, USA.

Nuclear factor-κB (NF-κB) regulates cellular responses to inflammation and aging, and alterations in NF-κB signaling underlie the pathogenesis of multiple human diseases. Effective clinical therapeutics targeting this pathway remain unavailable. In primary human keratinocytes, we found that hypochlorite (HOCl) reversibly inhibited the expression of CCL2 and SOD2, two NF-κB–dependent genes. In cultured cells, HOCl inhibited the activity of inhibitor of NF-κB kinase (IKK), a key regulator of NF-κB activation, by oxidizing cysteine residues Cys114 and Cys115. In NF-κB reporter mice, topical HOCl reduced LPS-induced NF-κB signaling in skin. We further evaluated topical HOCl use in two mouse models of NF-κB–driven epidermal disease. For mice with acute radiation dermatitis, topical HOCl inhibited the expression of NF-κB–dependent genes, decreased disease severity, and prevented skin ulceration. In aged mice, topical HOCl attenuated age-dependent production of p16INK4a and expression of the DNA repair gene Rad50. Additionally, skin of aged HOCl-treated mice acquired enhanced epidermal thickness and proliferation, comparable to skin in juvenile animals. These data suggest that topical HOCl reduces NF-κB–mediated epidermal pathology in radiation dermatitis and skin aging through IKK modulation and motivate the exploration of HOCl use for clinical aims.

Biofilm in Diabetic Foot Ulcers: A Systematic Narrative Review

George Theodorakopoulos^1,2,3,4,5,6 | David G. Armstrong^7,8,9

¹Wound and Foot Specialist, Athens, Greece | ²Technological Educational Institute of Patras, Patras, Greece | ³Queen Margaret University, Edinburgh, UK | ⁴University of West Attica, Aigaleo, Greece | ⁵University of Thessaly, Larisa, Greece | ⁶National and Kapodistrian University of Athens, Athens, Greece | ⁷Southwestern Academic Limb Salvage Alliance (SALSA), Los Angeles, California, USA | ⁸Center to Stream Healthcare in Place (C2SHiP), Alexandria, Virginia, USA | ⁹Keck School of Medicine, University of Southern California, Los Angeles, California, USA

Correspondence: George Theodorakopoulos (该Email地址已收到反垃圾邮件插件保护。要显示它您需要在浏览器中启用JavaScript。)

Received: 27 June 2025 | Revised: 29 October 2025 | Accepted: 6 November 2025

ABSTRACT

Biofilms are a key driver of chronicity and treatment failure in diabetic foot ulcers (DFUs), yet clinical evidence quantifying their impact and management remains fragmented. This systematic narrative review synthesised recent evidence (2015–2025) on the prevalence, diagnostics, and management of biofilm in DFUs. A Systematic Review of the Literature (SRL) was conducted following PRISMA 2020 guidelines across PubMed/MEDLINE, Scopus, Cochrane Library and ScienceDirect. Eligible studies included adults with DFUs reporting biofilm/bioburden metrics or interventions aimed at biofilm disruption. Risk of bias was assessed using RoB 2 for randomised trials and ROBINS-I for non-randomised studies. Data were narratively synthesised by evidence tier (Tier 1=clinical; Tier 2=preclinical/mechanistic). Of 600 records screened, 25 studies met inclusion criteria (Tier 1 n=9; Tier 2 n=5; reviews n=11). Over half of bacterial isolates in DFUs were biofilm producers, with multidrug resistance exceeding 90% in several cohorts. Fungi were detected in 31% of ulcers by qPCR but only 9% by culture. Tier 1 clinical evidence supports standard care components—debridement, antiseptics, and negative-pressure wound therapy—for improved healing, though direct anti-biofilm outcomes remain limited. Emerging strategies (enzymatic agents, peptides, cold plasma, smart dressings) show promise in vitro but lack clinical translation. Evidence for direct antibiofilm efficacy in DFUs remains scarce. Current data justify maintaining guideline-based care while prioritising trials that integrate validated biofilm endpoints, standardised microbiological methods, and antifungal components. Distinguishing established from experimental approaches is essential to advancing safe, evidence-based biofilm management in DFUs.

Role of an acidic environment in the treatment of diabetic foot infections: A review

Basavraj Nagoba, Ajay Gavkare, Abhijit Rayate, Sachin Mumbre, Arunkumar Rao, Basavraj Warad, Neeta Nanaware, Nawab Jamadar

Basavraj Nagoba, Department of Microbiology, MIMSR Medical College, Latur 413512, Maharashtra, India

Ajay Gavkare, Department of Physiology, MIMSR Medical College, Latur 413512, Maharashtra, India

Abhijit Rayate, Basavraj Warad, Department of Surgery, MIMSR Medical College, Latur 413512, Maharashtra, India

Sachin Mumbre, Department of Community Medicine, Ashwini Rural Medical College, Solapur 413001, Maharashtra, India

Arunkumar Rao, Department of Orthopedics, MIMSR Medical College, Latur 413512, India

Neeta Nanaware, Department of Physiology, Government Medical College, Latur 413512, Maharashtra, India

Nawab Jamadar, Department of Anesthesiology, MIMSR Medical College, Latur 413512, Maharashtra, India

Corresponding author: Basavraj Nagoba, PhD, Assistant Dean, Research, Professor, Department of Microbiology, MIMSR Medical College, Vishwanathpuram, Ambejogai Road, Latur 413512, Maharashtra, India. 该Email地址已收到反垃圾邮件插件保护。要显示它您需要在浏览器中启用JavaScript。

Abstract

Management of diabetic foot ulcers is the biggest challenge to the clinician, as conventional antibiotic therapies and local wound care have their own limitations. They are not effective for control of infections and promotion of healing because of cytotoxic effects. In view of cytotoxicity of routinely used topical antiseptic agents, this article focuses on the search of an ideal topical antiseptic agent that is safe and effective in controlling infectious agents and also in promoting the healing process. This review focuses on the use of various acids such as citric, acetic, hyaluronic, and hypochlorous acids as topical agents in diabetic foot infections. This article also focuses on the different roles of acids in the treatment of diabetic foot infections.

Key Words: Diabetic foot ulcer; Infection; Management; Topical agents; Acids; Role of acids

Core Tip: Diabetic foot ulcer is the most serious complication of diabetes mellitus. The biggest challenge is to find an ideal topical antiseptic agent that is safe and effective in controlling infectious agents and promoting the healing process. This article focuses on

the use of acids as topical agents to control diabetic foot infections, with special emphasis on the different roles of citric, acetic, hyaluronic, and hypochlorous acids in the effective management of diabetic foot ulcers.

Citation: Nagoba B, Gavkare A, Rayate A, Mumbre S, Rao A, Warad B, Nanaware N, Jamadar N. Role of an acidic environment in the treatment of diabetic foot infections: A review. World J Diabetes 2021; 12(9): 1539-1549