• 한국산학기술학회논문지
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• 제18권1호
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• pp.295-301
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• 2017

보툴리눔 신경독소는 콜린성 신경말단(부)을 선택적으로 공격하여 신경마비를 일으키는 신경독소로서, 그람양성을 띠고 내성포자를 형성하는 절대혐기성 세균인 보툴리눔 균(Clostridium botulinum)이 만들어낸다. 이 중 보툴리눔 A형 독소(BoNT/A)는 음식물과 물을 오염시킬 수 있으며 생물 무기나 생물 협박물질로 사용될 수도 있다. 이 때문에 독성을 탐지할 수 있는 예민한 분석방법과 중독을 치료할 수 있는 효능 있는 항독소를 개발해낼 필요성이 제기되어 왔다. 본 연구에서는 BoNT/A를 중화할 수 있는 단일클론 항체(mAb)를 생산하기 위하여 BoNT/A로 면역된 토끼의 항혈청에서 유래한 scFv 라이브러리를 인간 IgG와 융합시켰다. 그렇게 재조합된 scFvIgG 항체 단백질을 안정된 세포주에서 발현시켰고 항체 친화 크로마토그래피를 사용하여 scFvIgG mAb 단백질을 정제하였다. ELISA로 정제된 scFvIgG mAb 단백질의 효율성을 확인하였고, in vivo 실험으로 BoNT/A에 대한 중화능을 시험하였다. 독성 중화능 실험은 마우스를 사용하여 수행하였는데, 그 결과 scFvIgG 항체(10 ug)는 BoNT/A(100,000 $LD_{50}$)의 독성이 주입된 마우스를 완전히 방어하지는 못하지만 마우스의 생존 기간을 현격하게 연장시키는 것이 확인되었다. 이러한 결과들은 이 scFvIgG mAb가 BoNT/A를 중화하는 효능을 가지고 있다는 점을 제시한다.

• The Korean Journal of Physiology and Pharmacology
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• 제19권4호
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• pp.349-355
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• 2015

We examined the effects of peripherally or centrally administered botulinum neurotoxin type A (BoNT-A) on orofacial inflammatory pain to evaluate the antinociceptive effect of BoNT-A and its underlying mechanisms. The experiments were carried out on male Sprague-Dawley rats. Subcutaneous (3 U/kg) or intracisternal (0.3 or 1 U/kg) administration of BoNT-A significantly inhibited the formalin-induced nociceptive response in the second phase. Both subcutaneous (1 or 3 U/kg) and intracisternal (0.3 or 1 U/kg) injection of BoNT-A increased the latency of head withdrawal response in the complete Freund's adjuvant (CFA)-treated rats. Intracisternal administration of N-methyl-D-aspartate (NMDA) evoked nociceptive behavior via the activation of trigeminal neurons, which was attenuated by the subcutaneous or intracisternal injection of BoNT-A. Intracisternal injection of NMDA up-regulated c-Fos expression in the trigeminal neurons of the medullary dorsal horn. Subcutaneous (3 U/kg) or intracisternal (1 U/kg) administration of BoNT-A significantly reduced the number of c-Fos immunoreactive neurons in the NMDA-treated rats. These results suggest that the central antinociceptive effects the peripherally or centrally administered BoNT-A are mediated by transcytosed BoNT-A or direct inhibition of trigeminal neurons. Our data suggest that central targets of BoNT-A might provide a new therapeutic tool for the treatment of orofacial chronic pain conditions.

• The Korean Journal of Pain
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• 제18권1호
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• pp.29-33
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• 2005

Background: Chronic headache (CH) constitutes a significant public health problem, impacting on both the individual sufferer and society. Patients with CH, unresponsive to drug therapy or nerve block, suffer considerable disability due to the frequency and severity of attacks; therefore, they should be considered for novel therapy. Botulinum toxin type A (BoNT-A) has shown significant promise in the management of CH. In this paper, we review recent evidence on the efficacy of BoNT-A, and also report our experience with this treatment in CH patients. Methods: BoNT-A was used to treat 69 CH patients, including 47 in a chronic migraine group and 22 in a non-migraine CH group, who showed therapy-resistance to palliative drug or nerve block. We investigated the demography, dosage and site of BoNT-A injection, and used a visual analogue scale (VAS) for pain and the degree of satisfaction. The data were analyzed using t-tests and a Friedman repeated measures analysis of variance on ranks. Results: Significant decreases in the VAS for pain were found in both the chronic migraine and non-migraine CH groups, from 2, 4 and 12 weeks and from 4 and 12 weeks, respectively, after BoNT-A administration (P < 0.05). The chronic migraine group showed significantly lower VAS scores for pain than the non-migraine CH group from 2, 4 and 12 weeks after the BoNT-A administration (P < 0.05). Twenty eight patients (59.2%) in the chronic migraine group and eight (36.4%) in the non-migraine CH were satisfied with the BoNT-A treatment. Conclusions: This clinical study revealed that the use of BoNT-A demonstrated efficacy for CH patients resistant to drug therapy or nerve block. Moreover, BoNT-A proved itself more effective in the chronic migraine than non-migraine CH group.

• 한국임상수의학회지
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• 제28권3호
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• pp.273-279
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• 2011

보툴리늄 톡신(BoNT/A)은 사람에서 안전하고 효과적인 주름치료제로 적용되고 있으며 최근에는 주름치료효과 이외의 효능에 관한 연구가 활발히 진행되고 있다. 본 연구의 목적은 개 피부에 히스타민을 피하 주입하여 소양증을 유발한 다음 보툴리늄 톡신의 항소양 효과를 평가하는 것이다. 총 5 마리의 비글을 이용하여 우측 배측 흉부 피부에 보툴리늄 톡신 0.05 ml (5unit)를 주입한 처치부위와 좌측 배측 흉부 피부에 0.05 ml의 생리식염수를 주입한 대조부위를 비교하였다. 보툴리늄 톡신 투여 전, 투여 후 1, 3, 7일에 Histamine 을 피내주입하여 소양증을 유발하였다. 소양증의 정도, 팽진의 지름과 두께, 홍반수치 및 피부표면 온도를 측정하여 보툴리늄 톡신 주입 효과를 평가하였다. 소양증의 정도는 처치부위에서 유의하게 감소하였으며(p < 0.05) 팽진의 지름과 두께도 처치부위에서 유의하게 감소했다(p < 0.05). 홍반수치는 최초 히스타민을 피내투여한 직후에 처치부위와 대조부위에서 모두 증가했으나 대조부위에 비해서 처치부위에서 적게 증가하였다. 피부표면온도는 처치부위에서 유의하게 감소하였다(p < 0.05). 본 연구결과 보툴리늄 톡신은 개 피부에서 히스타민에 의한 소양증에 대한 항소양 효과를 보였으며 임상적으로 극심한 국소 소양증을 보이는 피부질환에 대해 적용할 수 있을 것으로 사료된다.

• 대한후두음성언어의학회지
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• 제30권2호
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• pp.77-81
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• 2019

Dysphagia may result from dysfunction of any of the components involved in the complex neuromuscular interaction of swallowing. Hyperfunction of any of the muscles involved in swallowing is a frequent cause of dysphagia. The cricopharyngeus muscle (CPM) is a key component of the upper esophageal sphincter. Cricopharyngeus muscle dysfunction (CPD) refers to the muscle's failure to appropriately and completely relax or expand during deglutition. A variety of disease processes may cause CPD and accurate diagnosis is paramount for appropriate treatment. In appropriately selected patients, intervention at the CPM may yield significant improvement in dysphagia. Interventions include nonsurgical, pharyngoesophageal segment dilatation, botulinum toxin (BoNT) injection, and criccopharyngeal myotomy. Injections of BoNT in patients with CPD have been reported to result in marked relief of dysphagia. Different techniques for instilling BoNT into the CPM have been described. Awake, in-office CPM BoNT injection with electromyography and/or fluoroscopic or ultrasound guidance is performed transcervically or via flexible endoscopy. Operative CPM BoNT injection involves rigid laryngoscopy and esophagoscopy with direct visualization of the CPM. BoNT should be prepared in low-volume, high-concentration dilutions to minimize the potential for undesired diffusion of the toxin. The effects of BoNT occur within weeks of injection and typically last up to 5 or 6 months.

• The Korean Journal of Physiology and Pharmacology
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• 제22권5호
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• pp.539-546
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• 2018

Botulinum toxin type A (BoNT/A) has been used therapeutically for various conditions including dystonia, cerebral palsy, wrinkle, hyperhidrosis and pain control. The substantia gelatinosa (SG) neurons of the trigeminal subnucleus caudalis (Vc) receive orofacial nociceptive information from primary afferents and transmit the information to higher brain center. Although many studies have shown the analgesic effects of BoNT/A, the effects of BoNT/A at the central nervous system and the action mechanism are not well understood. Therefore, the effects of BoNT/A on the spontaneous postsynaptic currents (sPSCs) in the SG neurons were investigated. In whole cell voltage clamp mode, the frequency of sPSCs was increased in 18 (37.5%) neurons, decreased in 5 (10.4%) neurons and not affected in 25 (52.1%) of 48 neurons tested by BoNT/A (3 nM). Similar proportions of frequency variation of sPSCs were observed in 1 and 10 nM BoNT/A and no significant differences were observed in the relative mean frequencies of sPSCs among 1-10 nM BoNT/A. BoNT/A-induced frequency increase of sPSCs was not affected by pretreated tetrodotoxin ($0.5{\mu}M$). In addition, the frequency of sIPSCs in the presence of CNQX ($10{\mu}M$) and AP5 ($20{\mu}M$) was increased in 10 (53%) neurons, decreased in 1 (5%) neuron and not affected in 8 (42%) of 19 neurons tested by BoNT/A (3 nM). These results demonstrate that BoNT/A increases the frequency of sIPSCs on SG neurons of the Vc at least partly and can provide an evidence for rapid action of BoNT/A at the central nervous system.

• 대한두개안면성형외과학회지
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• 제22권1호
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• pp.1-10
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• 2021

Botulinum toxin type A (BoNT-A), onabotulinumtoxinA (Botox) was approved by the United States Food and Drug Administration for temporary improvement of glabellar lines in patients 65 years and younger in 2002, and has also been used widely for aesthetic purposes such as hyperhidrosis, body shape contouring, and other noninvasive facial procedures. BoNT-A inhibits presynaptic exocytosis of acetylcholine (ACh)-containing vesicles into the neuromuscular junction at cholinergic nerve endings of the peripheral nervous system, thereby paralyzing skeletal muscles. ACh is the most broadly used neurotransmitter in the somatic nervous system, preganglionic and postganglionic fibers of parasympathetic nerves, and preganglionic fibers or postganglionic sudomotor nerves of sympathetic nerves. The scientific basis for using BoNT-A in various cosmetic procedures is that its function goes beyond the dual role of muscle paralysis and neuromodulation by inhibiting the secretion of ACh. Although the major target organs for aesthetic procedures are facial expression muscles, skeletal body muscles, salivary glands, and sweat glands, which are innervated by the somatic or autonomic nerves of the peripheral cholinergic nerve system, few studies have attempted to directly explain the anatomy of the areas targeted for injection by addressing the neural physiology and rationale for specific aesthetic applications of BoNT-A therapy. In this article, we classify the various cosmetic uses of BoNT-A according to the relevant component of the peripheral nervous system, and describe scientific theories regarding the anatomy and physiology of the cholinergic nervous system. We also review critical physiological factors and conditions influencing the efficacy of BoNT-A for the rational aesthetic use of BoNT-A. We hope that this comprehensive review helps promote management policies to support long-term, safe, successful practice. Furthermore, based on this, we look forward to developing and expanding new advanced indications for the aesthetic use of BoNT-A in the future.

• The Korean Journal of Pain
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• 제24권3호
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• pp.164-168
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• 2011

Obturator nerve block has been commonly used for pain management to prevent involuntary reflex of the adductor thigh muscles. One of several options for this block is chemical neurolysis. Neurolysis is done with chemical agents. Chemical agents used in the neurolysis of the obturator nerve have been alcohol, phenol, and botulinum toxin. In the current case, a patient with spasticity of the adductor thigh muscle due to cervical cord injury had obturator nerve neurolysis done with botulinum toxin type B (BoNT-B). Most of the previous studies have used BoNT-A with only a few reports that have used BoNT-B. BoNT-B has several advantages and disadvantages over BoNT-A. Thus, we report herein a patient who successfully received obturator nerve neurolysis using BoNT-B to treat adductor thigh muscle spasm.

• BMB Reports
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• 제39권5호
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• pp.642-647
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• 2006

Botulinum neurotoxin A (BoNT/A) has been used therapeutically to treat muscular hypercontractions and sudomotor hyperactivity and it has been reported that BoNT/A might have analgesic properties in headache. PEP-1 peptide is a known carrier peptide that delivers fulll-ength native proteins in vitro and in vivo. In this study, a BoNT/A gene were fused with PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame PEP-1-BoNT/A fusion protein. The expressed and purified PEP-1-BoNT/A fusion proteins were efficiently transduced into cells in a time- and dose-dependent manner when added exogenously in a culture medium. In addition, immuno-histochemical analysis revealed that PEP-1-BoNT/A fusion protein efficiently penetrated into the epidermis as well as the dermis of the subcutaneous layer, when sprayed on mice skin. These results suggest that PEP-1-BoNT/A fusion protein provide an efficient strategy for therapeutic delivery in various human diseases related to this protein.

• Anatomy and Cell Biology
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• 제56권3호
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• pp.293-298
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• 2023

The mentalis muscle is a paired muscle originating from the alveolar bone of the mandible. This muscle is the main target muscle for botulinum neurotoxin (BoNT) injection therapy, which aims to treat cobblestone chin caused by mentalis hyperactivity. However, a lack of knowledge on the anatomy of the mentalis muscle and the properties of BoNT can lead to side effects, such as mouth closure insufficiency and smile asymmetry due to ptosis of the lower lip after BoNT injection procedures. Therefore, we have reviewed the anatomical properties associated with BoNT injection into the mentalis muscle. An up-to-date understanding of the localization of the BoNT injection point according to mandibular anatomy leads to better injection localization into the mentalis muscle. Optimal injection sites have been provided for the mentalis muscle and a proper injection technique has been described. We have suggested optimal injection sites based on the external anatomical landmarks of the mandible. The aim of these guidelines is to maximize the effects of BoNT therapy by minimizing the deleterious effects, which can be very useful in clinical settings.