Multiroute administration of Wharton’s jelly mesenchymal stem cells in chronic complete spinal cord injury: A phase I safety and feasibility study

Table 1 Enrollment criteria and assessment timeline

Criteria	Details
Inclusion criteria	Age ≥ 18 years
	Radiologically confirmed chronic SCI
	No chronic illnesses (e.g., cancer, organ failure)
	Life expectancy > 12 months
	Severe disability (wheelchair-bound, assistance required)
	No significant functional improvement for at least 3 months
	Commitment to standard care and follow-up
	Provide written informed consent
Exclusion criteria	Significant medical/psychiatric conditions affecting safety
	Recent severe infections (meningitis, sepsis, etc.)
	Uncontrolled seizure disorder
	CNS neoplasm or cancer history (last 5 years)
	Abnormal blood cell counts (WBC ≥ 15000/μL, platelets ≤ 100000/μL)
	Abnormal liver/kidney function (AST/ALT > 3 × ULN, creatinine > 1.5 × ULN)
	Prior participation in other investigational stem cell trials
	Patient decision to withdraw or death during study
Treatment timeline	Baseline assessment (week 0): Neurological and functional evaluations
	MSC therapy initiation (week 1-8): 4 rounds of triple-route administration
	Short-term follow-up (weeks 1-12): Safety and early functional outcomes
	Mid-term assessment (month 6): Progress evaluation
	Long-term follow-up (month 12): Final outcome assessment

SCI: Spinal cord injury; CNS: Central nervous system; AST: Aspartate aminotransferase; ALT: Alanine aminotransferase; ULN: Upper limit of normal; MSC: Mesenchymal stem cell.

Table 2 Study population

Characteristic		n	%
Age, years	19	1	16.7
	20	1	16.7
	29	3	50.0
	39	1	16.7
Sex	Female	1	16.7
	Male	5	83.3
Cause of SCI	Fell from tree	1	16.7
	Firearm injury	2	33.3
	Ski accident	1	16.7
	Traffic accident	2	33.3
Level of injury	T11 burst fracture	1	16.7
	T12 burst fracture with T11-T12 and T12-L1 traumatic disc herniations	1	16.7
	T12 burst fracture with dislocation	1	16.7
	T5 burst fracture	1	16.7
	T7-8 burst fracture	1	16.7
	T8-9 burst fracture	1	16.7
Pre-transplantation treatments	Only physiotherapy	3	50.0
	T11 laminectomy + physiotherapy	1	16.7
	T12 corpectomy + posterior instrumentation + laminectomy + physiotherapy	1	16.7
	T12 corpectomy + posterior instrumentation + laminectomy + physiotherapy	1	16.7
Comorbidity	None	6	100.0
Duration of SCI and first transplantation	6 months	1	16.7
	9 months	2	33.3
	12 months	1	16.7
	4 years	1	16.7
	12 years	1	16.7

SCI: Spinal cord injury.

Table 3 Wharton’s jelly-derived mesenchymal stem cell quality control parameters

	QC parameters
Cell count and viability	Cell viability > 90. Cell count: Varies according to the patient
Immunophenotyping	CD90, CD44, CD105, CD73 ≥ 95%. CD45, CD34, CD19, CD11B, HLA-DR, CD14 ≤ 2% (Figure 1A)
Gene expression	SOX2, OCT4, TERT, REX1, NES, BMP2, RUNX2, PPARG, JAG1, ICAM1, VCAM1, VIM. It was concluded that there was strong positivity in genes with Ct values lower than 29, positive in genes with values between 30-37, and poor expression in samples with Ct values between 38-40 or there may be environmental contamination (Figure 1B)
Telomerase	The result should be lower than that of the positive control (Figure 1C)
Mycoplasma	Negative
Endotoxin	< 0.0625 EU/mL

HLA: Human leukocyte antigen; QC: Quality control.

Table 4 Administration schedule

Rounds	Route	WJ-MSC
Round 1	IT	1 × 106 cells/kg in 3 mL
	IV	1 × 106 cells/kg in 30 mL
	IM	1 × 106 cells/kg in 20 mL
Round 2 (week 1)	IT	1 × 106 cells/kg in 3 mL
	IV	1 × 106 cells/kg in 30 mL
	IM	1 × 106 cells/kg in 20 mL
Round 3 (week 4)	IT	1 × 106 cells/kg in 3 mL
	IV	1 × 106 cells/kg in 30 mL
	IM	1 × 106 cells/kg in 20 mL
Round 4 (week 6)	IT	1 × 106 cells/kg in 3 mL
	IV	1 × 106 cells/kg in 30 mL
	IM	1 × 106 cells/kg in 20 mL

WJ-MSC: Wharton’s jelly derived mesenchymal stem cells; IT: Intrathecal; IM: Intramuscular; IV: Intravenous.

Table 5 Results of the Friedman test for changes in Functional Independence Measure Motor score values before and after intervention

	n	Mean	SD	Mean rank	χ2	df	P value
Preoperative	6	42.50	2.26	1.50
Postoperative week 1	6	43.00	1.90	1.67
Postoperative month 1	6	45.67	1.97	2.83	29.505	5	0.000
Postoperative month 2	6	48.00	1.90	4.00
Postoperative month 6	6	50.50	2.17	5.00
Postoperative month 12	6	56.17	2.99	6.00

SD: Standard deviation.

Table 6 Results of the Friedman test for changes in the Modified Ashworth scale (right values) before and after intervention

	n	Mean	SD	Mean rank	χ2	df	P value
Preoperative	6	8.00	2.45	5.83
Postoperative week 1	6	7.17	3.06	5.17
Postoperative month 1	6	5.33	2.66	3.67	28.452	5	0.000
Postoperative month 2	6	3.83	2.48	3.00
Postoperative month 6	6	3.00	2.19	2.08
Postoperative month 12	6	2.17	1.94	1.25

SD: Standard deviation.

Table 7 Results of the Friedman test for changes in the Modified Ashworth scale (left values) before and after intervention

	n	Mean	SD	Mean rank	χ2	df	P value
Preoperative	6	7.83	2.40	5.75
Postoperative week 1	6	7.17	2.64	5.25
Postoperative month 1	6	5.50	2.43	3.92	29.030	5	0.000
Postoperative month 2	6	4.17	2.64	2.92
Postoperative month 6	6	3.00	2.28	1.92
Postoperative month 12	6	2.17	1.84	1.25

SD: Standard deviation.

Table 8 Results of the Friedman test for changes in the American Spinal Injury Association Motor score values before and after intervention

	n	Mean	SD	Mean rank	χ2	df	P value
Preoperative	6	1.67	2.66	2.00
Postoperative week 1	6	1.67	2.66	2.00
Postoperative month 1	6	3.33	3.93	2.75	23.938	5	0.000
Postoperative month 2	6	7.67	5.28	4.08
Postoperative month 6	6	12.00	8.10	4.75
Postoperative month 12	6	15.00	11.23	5.42

SD: Standard deviation.

Table 9 Results of the Friedman test for changes in the American Spinal Injury Association Sensory score (Light Touch values) before and after intervention

	n	Mean	SD	Mean rank	χ2	df	P value
Preoperative	6	62.00	18.42	1.50
Postoperative week 1	6	65.33	18.23	1.83
Postoperative month 1	6	70.33	18.26	2.75	28.299	5	0.000
Postoperative month 2	6	74.67	19.04	4.08
Postoperative month 6	6	78.67	20.73	5.08
Postoperative month 12	6	81.67	22.50	5.75

SD: Standard deviation.

Table 10 Results of the Friedman test for changes in the American Spinal Injury Association Sensory Score (Pin-Prick values) before and after intervention

	n	Mean	SD	Mean rank	χ2	df	P value
Preoperative	6	62.00	18.42	1.50
Postoperative week 1	6	63.67	18.13	1.83	28.838	5	0.000
Postoperative month 1	6	68.67	17.47	2.75
Postoperative month 2	6	74.33	19.82	3.92
Postoperative month 6	6	78.33	20.92	5.08
Postoperative month 12	6	81.33	22.26	5.92

SD: Standard deviation.

Table 11 Results of the Wilcoxon signed-rank test for changes in the Wexner Incontinence Score values before and after intervention

Wexner Incontinence Score	Ranks	n	Mean rank	Sum of ranks	Z score	P value
Preoperative	Negative ranks	0	3.50	21.00	-2.226	0.026
	Positive ranks	6	0.00	0.00
Postoperative	Ties	0
	Total	6

Table 12 Results of the Wilcoxon signed-rank test for changes in the Qualiveen Short Form values before and after intervention

SFQ	Ranks	n	Mean rank	Sum of ranks	Z score	P value
Preoperative	Negative ranks	0	3.50	21.00	-2.214	0.027
	Positive ranks	6	0.00	0.00
Postoperative	Ties	0
	Total	6

SFQ: Qualiveen Short Form.

Table 13 Early and late complications

Complications		Patient No. 11				Patient No. 21				Patient No. 31				Patient No. 41				Patient No. 51				Patient No. 61
		1st	2nd	3rd	4th	1st	2nd	3rd	4th	1st	2nd	3rd	4th	1st	2nd	3rd	4th	1st	2nd	3rd	4th	1st	2nd	3rd	4th
Early	Infection	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Fever	-	+	+	-	-	-	-	-	+	+	-	-	-	-	-	-	-	+	-	-	+	-	+	-
	Pain	-	-	-	-	-	-	-	-	+	+	+	-	-	-	-	-	+	-	-	-	-	-	-	-
	Headache	-	-	-	-	-	-	-	-	-	-	+	-	-	-	-	-	-	-	-	-	+	+	+	-
	Increased C-reactive protein levels	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Leukocytosis	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Allergic reaction or shock	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Perioperative complications	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Late	Secondary infections	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Urinary tract infections	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Deterioration of neurological status	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Neuropathic pain	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Carcinogenesis	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-

¹Administration.