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BoNT-A

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BoNT-A is a highly purified form of the Botulinum Neurotoxin Type A. It is a biological product used in research applications. BoNT-A functions by inhibiting neurotransmitter release at the neuromuscular junction.

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18 protocols using BoNT-A

The animals used in this study were maintained in accordance with the Guide for the Care and Use of Laboratory Animals published by the US National Institution of Health (NIH Publication No 85-23, revised 1996) and the Policy of Animal Care and Use Committee of Tongji University. Male Sprague-Dawley rats (200–220 g) and rats at the 1st, 4th, and 8th week after birth were obtained from B&K Universal Group Limited (Shanghai, China). Adult rats were fed with chow and water ad libitum at the Animal Center of Tongji Hospital, and maintained under controlled temperature (20–22°C) and a 12 h light/dark cycle.
Adult rats were randomly divided into three groups: control group (n = 21), BoNT/A group (n = 21), and agrin-Ab groups. BoNT/A (BOTOX®, Allergan, Co. Mayo, Ireland) was reconstituted in saline (NS) to a final concentration of 2 U/100 μL. Animals of the BoNT/A and agrin-Ab groups were injected unilaterally with 100 μL BoNT/A in the right gastrocnemius muscle under anesthesia with an intraperitoneal injection of pentobarbital (30 mg/kg). On the 3rd day after BoNT/A injection, the each subgroups of agrin-Ab were injected with 100 μL agrin-Ab (R&D Systems, Minnesota, CA, United States) at a dosage of 0.6, 2, 6, 20, or 60 μg respectively at once. Controls received an equivalent volume of NS injections in the right gastrocnemius muscle.
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BoNT-A (100 U; Botox, Allergan, Irvine, California, CA, USA) was reconstituted with 1 mL of non-preserved, sterile, isotonic 0.9% saline at room-temperature. A total of 80 U of BoNT-A was used for each patient, divided into 30 U for each masseter muscle and 10 U for each anterior temporalis muscle23 (link). A total of 0.8 mL of saline solution served as a control. Bilateral intramuscular injections were performed using a 1 mL syringe with a 30-gauge needle. Subjects were asked to clench their teeth to delineate the muscle area to be injected (masseter and anterior temporalis). After careful aspiration, a total of 5 injections per muscle were applied 5 mm apart. If the aspiration was positive, the needle was moved slightly and aspiration was repeated until it was negative. The patients and the researcher applying the injections were blinded to the treatment. Another researcher not present during injections, opened the sealed envelope, reconstituted BoNT-A and prepared the syringes according to the subjects distribution. Injections were applied in a single session.
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3

Standardized Botulinum Toxin Injections for Facial Aesthetics

BoNT-A (100 U, Botox®, Allergan, Irvine, CA, USA) was reconstituted with 1.0 mL of preservative-free saline solution so that each 0.1 mL corresponded to 10 U of product. The doses applied in the frontalis, procerus, and corrugator supercili muscles followed the Italian consensus for onabotulinumtoxinA (Botox®) injections [10 (link)]. Therefore, 12 U, divided into 6 injection points, were applied in the frontalis muscle, a single 4-U injection was applied in the procerus muscle, and single 4U and 3U injections were applied, respectively, in the medial head and lateral part of the corrugator supercili muscle. The injected doses were not adjusted for individual esthetic needs. The injections were performed by the same researcher, who did not know the corresponding group of the patient and was not involved in any other process of this study. Injections were performed in a single session, and no compensatory injections were made in the case of not reaching an optimal result.
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In the control group, patients received routine rehabilitation treatment, including oral anti-spasticity medications and the use of ankle-foot orthotics. In this study, we did not intervene in the choice of medication, method of rehabilitation, and brace type. In the experimental group, patients received BoNT-A (Allergan) injection and routine rehabilitation treatment without oral anti-spasticity medications. The injection was administered by the same physician using electrical stimulation-guided injection. The injection dose and target sites were selected by the physician after individual assessment, referring to Chinese guidelines for the treatment of adult limb spasm with BoNT (Li et al., 2015 (link)). The main target muscles were selected mainly depending on gait patterns including quadriceps femoris (QF), gastrocnemius (GS), tibialis posterior (TP), flexor hallucis longus (FHL), FDL, FD brevis (FDB), and FH brevis (FHB) [specific operations followed table (Table 1)].
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5

Submandibular Gland Botulinum Toxin Injection and Duct Ligation

Both BoNT‐A injections and bilateral submandibular duct ligation were performed in an outpatient setting. BoNT‐A (Allergan, Nieuwegein, the Netherlands) was administered to the submandibular glands under general anaesthesia, fractioning 25U in 0.9% saline over each submandibular gland using a 25‐gauge needle and ultrasound guidance. Patients were treated under general anaesthesia and ultrasound guidance to decrease the risk of adverse events in the case of extra‐glandular BoNT‐A. In participants with previous BoNT‐A treatment, BoNT‐A injections or bilateral submandibular duct ligation were performed at least 6 months after the last injection to prevent a carry‐over effect.
Bilateral submandibular duct ligation was also performed under general anaesthesia. The submandibular ducts were traced, dissected, and ligated, applying two metal vascular clips to each duct. Intraoral absorbable sutures closed the incision.6 All patients who had received bilateral submandibular duct ligation received amoxicillin/clavulanic acid and paracetamol plus diclofenac for 7 and 5 days respectively.
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Bilateral injections into the submandibular glands with BoNT‐A (Botox; Allergan) were performed by the same physician (PJ) using ultrasound guidance and general anaesthesia. Botox was diluted in 0.9% saline solution (25U/ml) and 1ml was administered over two or three sites throughout the gland using a Spinocan needle (25 Ga; B. Braun Medical B.V., Oss, the Netherlands).
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BoNT-A was administered 5 days after cyclophosphamide injection to fully induce the OAB symptoms. The rats were anesthetized with 10 mg/kg of Zoletil 50® (Vibac Laboratories, Carros, France) before BoNT-A administration. An incision was made in the skin on the lateral surface of the thigh and the biceps femoris muscle was cut across, exposing the sciatic nerve and its three terminal branches: the sural, common peroneal, and tibial nerves. Then, using a tuberculin syringe fitted with a 30-gauge needle, 100 units of BoNT-A (Allergan TM, Mayo, Ireland) were placed directly onto the exposed nerve and were allowed to flow onto the surrounding fascia to mimic the distributive behaviour of the toxin after injection at the target sites.
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BoNT-A (100 U; Botox, Allergan, Irvine, California, CA, USA) was reconstituted using non-preserved sterile saline solution 0.9%. A single bilateral injection was applied in the masseter and anterior temporalis muscles using 30U and 10U of BoNT-A, respectively15 (link) by a calibrated researcher, distributed in five sites for each muscle. It was used a 1 mL syringe with a 30-gauge and 13 mm needle. Briefly, for the masseter, the injected sites were in the inferior part of the muscle (on mandibular angle), 5 mm apart from each other. For the anterior temporal muscles, sites were determined according to the functional test, considering the most prominent part, and must be 1 cm external to the eyebrow and 5 mm apart among them (Figure 2). Injections were performed bilaterally regardless the patient had pain in only one side of the face.
A, marked points for <a class="product" href="/product/KyPhCZIBPBHhf-iFxpw-/" target="_blank">BoNT-A</a> injection. B, temporal muscle injection; C, masseter muscle injectionThis injection technique consisted in inserting the needle into the soft tissue until reaching the bone; then, the needle was slightly moved to place the tip inside the muscle. Before injection, a careful aspiration was performed to avoid a possible intravascular administration.
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BoNT-A (100 U; Botox, Allergan, Irvine, California, CA, USA) was reconstituted using non-preserved sterile saline solution 0.9%. To blind the applications of BoNT-A and SS, each muscle received an injection containing 1 mL of the correspondent diluted BoNTA-L/M/H doses or SS by an investigator who was not involved in the dilution process. Doses of BoNT-A were based on previous reports [11 (link),12 (link),13 (link),14 (link)] and assigned according to group distribution (Figure 1). Bilateral intramuscular injections were performed using a 1-mL syringe with a 30-gauge needle. Subjects were asked to clench to delimit the muscle area (masseter and anterior temporalis) and a total of 5 injections per muscle with a separation of 5 mm were applied.
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After the bladder catheter was secured in place, the bladder was filled just enough to distend the bladder walls for ease of injection. Ten units of BoNT/A (onabotulinum toxin type A, Allergan Inc., Irvine, CA, USA) in 20μl of saline or 20μl of saline alone (sham procedure for controls) were injected in 5 equally divided doses, one in each wall and one in the dome using a 25μl Hamilton syringe.11 (link) Injection in the bladder tissues was confirmed visually by blebbing of the tissue at the injection site ensuring injection into all walls and the dome. We chose a dose of 10 units as this was above the minimal effective dose of 7.5 units demonstrated in a previous dose response study on female Sprague-Dawley rats.11 (link)
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